US20010010916A1 - Preparation of biopolymer arrays - Google Patents
Preparation of biopolymer arrays Download PDFInfo
- Publication number
- US20010010916A1 US20010010916A1 US09/820,476 US82047601A US2001010916A1 US 20010010916 A1 US20010010916 A1 US 20010010916A1 US 82047601 A US82047601 A US 82047601A US 2001010916 A1 US2001010916 A1 US 2001010916A1
- Authority
- US
- United States
- Prior art keywords
- head
- chamber
- substrate
- venturi
- fluid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/02—Burettes; Pipettes
- B01L3/0241—Drop counters; Drop formers
- B01L3/0268—Drop counters; Drop formers using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00277—Apparatus
- B01J2219/00351—Means for dispensing and evacuation of reagents
- B01J2219/0036—Nozzles
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00277—Apparatus
- B01J2219/00351—Means for dispensing and evacuation of reagents
- B01J2219/00378—Piezo-electric or ink jet dispensers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00277—Apparatus
- B01J2219/00497—Features relating to the solid phase supports
- B01J2219/00527—Sheets
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00585—Parallel processes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/0059—Sequential processes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00596—Solid-phase processes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/0061—The surface being organic
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00612—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports the surface being inorganic
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00614—Delimitation of the attachment areas
- B01J2219/00617—Delimitation of the attachment areas by chemical means
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00614—Delimitation of the attachment areas
- B01J2219/00617—Delimitation of the attachment areas by chemical means
- B01J2219/00619—Delimitation of the attachment areas by chemical means using hydrophilic or hydrophobic regions
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00623—Immobilisation or binding
- B01J2219/00626—Covalent
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
- B01J2219/00632—Introduction of reactive groups to the surface
- B01J2219/00637—Introduction of reactive groups to the surface by coating it with another layer
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00583—Features relative to the processes being carried out
- B01J2219/00603—Making arrays on substantially continuous surfaces
- B01J2219/00659—Two-dimensional arrays
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/0068—Means for controlling the apparatus of the process
- B01J2219/00686—Automatic
- B01J2219/00689—Automatic using computers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00718—Type of compounds synthesised
- B01J2219/0072—Organic compounds
- B01J2219/00722—Nucleotides
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00718—Type of compounds synthesised
- B01J2219/0072—Organic compounds
- B01J2219/00725—Peptides
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00718—Type of compounds synthesised
- B01J2219/0072—Organic compounds
- B01J2219/00729—Peptide nucleic acids [PNA]
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00718—Type of compounds synthesised
- B01J2219/0072—Organic compounds
- B01J2219/00731—Saccharides
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B40/00—Libraries per se, e.g. arrays, mixtures
- C40B40/04—Libraries containing only organic compounds
- C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B40/00—Libraries per se, e.g. arrays, mixtures
- C40B40/04—Libraries containing only organic compounds
- C40B40/10—Libraries containing peptides or polypeptides, or derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B40/00—Libraries per se, e.g. arrays, mixtures
- C40B40/04—Libraries containing only organic compounds
- C40B40/12—Libraries containing saccharides or polysaccharides, or derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B40/00—Libraries per se, e.g. arrays, mixtures
- C40B40/04—Libraries containing only organic compounds
- C40B40/14—Libraries containing macromolecular compounds and not covered by groups C40B40/06 - C40B40/12
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B60/00—Apparatus specially adapted for use in combinatorial chemistry or with libraries
- C40B60/14—Apparatus specially adapted for use in combinatorial chemistry or with libraries for creating libraries
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
- G01N2035/1027—General features of the devices
- G01N2035/1034—Transferring microquantities of liquid
- G01N2035/1041—Ink-jet like dispensers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
- G01N35/1065—Multiple transfer devices
- G01N35/1074—Multiple transfer devices arranged in a two-dimensional array
Definitions
- This invention relates to biopolymer arrays, particularly polynucleotide arrays such as DNA arrays, which are useful in diagnostic, screening, gene expression analysis, and other applications.
- Arrays of biopolymers such as arrays of peptides or polynucleotides (such as DNA or RNA), are known and are used, for example, as diagnostic or screening tools.
- Such arrays include regions (sometimes referenced as spots) of usually different sequence biopolymers arranged in a predetermined configuration on a substrate.
- the arrays when exposed to a sample, will exhibit a pattern of binding which is indicative of the presence and/or concentration of one or more components of the sample, such as an antigen in the case of a peptide array or a polynucleotide of particular sequence in the case of a polynucleotide array.
- the binding pattern can be detected, for example, by labeling all potential targets (for example, DNA) in the sample with a suitable label (such as a fluorescent compound), and accurately observing the fluorescence pattern on the array.
- Biopolymer arrays can be fabricated using in situ synthesis methods or deposition of the previously obtained biopolymers.
- the in situ synthesis methods include those described in U.S. Pat. No. 5,449,754 for synthesizing peptide arrays, as well as WO 98/41531 and the references cited therein for synthesizing polynucleotides (specifically, DNA).
- Such in situ synthesis methods can be basically regarded as repeating at each spot the sequence of: (a) deprotecting any previously deposited monomer so that it can now link with a subsequently deposited protected monomer; and (b) depositing a droplet of another protected monomer for linking.
- Different monomers may be deposited at different regions on the substrate during any one iteration so that the different regions of the completed array will have different desired biopolymer sequences.
- One or more intermediate further steps may be required in each iteration, such as oxidation, capping and washing steps.
- the deposition methods basically involve depositing biopolymers at predetermined locations on a substrate which are suitably activated such that the biopolymers can link thereto. Biopolymers of different sequence may be deposited at different regions of the substrate to yield the completed array. Washing or other additional steps may also be used. Reagents used in typical in situ synthesis are water sensitive, and thus the presence of moisture should be eliminated or at least minimized.
- Typical procedures known in the art for deposition DNA such as whole oligomers or cDNA, are to load a small volume of DNA in solution on the tip of a pin or in an open capillary and touch the pin or capillary to the surface of the substrate. When the fluid touches the surface, some of the fluid is transferred. The pin or capillary must be washed prior to picking up the next type of DNA for spotting onto the array. This process is repeated for many different sequences and, eventually, the desired array is formed.
- the DNA can be loaded into an inkjet head and fired onto the substrate.
- Such a technique has been described, for example, in PCT publications WO 95/25116 and WO 98/41531, and elsewhere. This method has the advantage of non-contact deposition.
- Still other methods include pipetting and positive displacement pumps such as the Bio-Dot A/D3000 Dispenser available from Bio-Dot Inc., Irvine, Calif., USA.
- the array sensitivity is dependent on having reproducible spots on the substrate. The location of each type of spot must be known and the spotted area should be uniformly coated with the DNA.
- the transfer device since DNA is expensive to produce, a minimum amount of the DNA solution should be loaded into any of the transfer mechanisms.
- any cross contamination of different DNA's must be lower than the sensitivity of the final array as used in a particular assay, to prevent false positive signals. Therefore, the transfer device must be easily cleaned after each type of DNA is deposited or the device must be inexpensive enough to be a disposable.
- the quantity of the assay sample is often limited, it is advantageous to make the spots small and closely spaced.
- inkjet deposition has advantages which include producing very small spot sizes. This allows high-density arrays to be fabricated. Furthermore, the spot size is uniform and reproducible as demonstrated by the successful use of inkjets in printers. Since it is a non-contact technique, ink-jet deposition will not scratch or damage the surface. Ink-jets have very high deposition rate, which facilitates rapid manufacture of arrays.
- an ink-jet deposition system used for fabricating a biopolymer array should meet a number of requirements. Specifically, the inkjet head must be capable of being loaded with very small volumes of DNA solution and function with minimal or no priming of the inkjets. The system should provide for easy purging of the working solution and readily flushed clean when required. When used for in-situ synthesis, the system should be able to to keep reagents isolated from moisture in the surrounding air.
- a negative backpressure that is, a pressure behind an orifice of the jet
- a pressure behind an orifice of the jet in the range of one to six inches of water
- Open-cell foam has been used to provide this negative backpressure in an inkjet printer in a manner disclosed in U.S. Pat. No. 4,771,295, such the capillarity of the foam creates the negative backpressure in an ink reservoir. While this is an easy and economical way to provide the required negative backpressure, the foam cannot be easily purged of the working fluid.
- a small rubber thimble similar to an eyedropper, can alternatively be used but the backpressure will vary as the reservoir is depleted.
- rubber is incompatible with the chemical reagents typically used in in-situ synthesis.
- a spring bag reservoir can be designed to control the backpressure of the fluid reservoir, however it requires a large working volume and is therefore not a good choice for the small reservoir volumes required by DNA or other biopolymer array fabrication.
- a regulated vacuum source could also be used. However, this may permit undesirable components, such as moisture, entering the head particularly during in situ synthesis. Additionally, purging the inkjet head then requires extra valves and a compressed nitrogen (or other suitable gas) source. Gravity is one of the easiest backpressure control means, however the backpressure changes as the fluid height drops and it requires too large a fluid volume to work properly for the small volumes encountered in an inkjet.
- the present invention provides a method of fabricating an array of different or the same moieties (for example, multiple different chemical compounds) on a substrate using one or more suitable fluids, and using a fluid dispensing head.
- the invention is particularly useful for the in situ process since it provides the required head pressure while facilitating isolation of reagents from moisture or other undesirable components.
- the invention is also applicable to the direct deposition of polynucleotides.
- the invention provides a method of fabricating an array of biopolymers using a biopolymer containing fluid, or one or more fluids containing a biomonomer.
- the head has at least one jet which can dispense droplets of a fluid onto a substrate, the jet including a chamber with an orifice, and including an ejector which, when activated, causes a droplet to be ejected from the orifice.
- the head may particularly be of a type commonly used in inkjet printers, in which a plurality of pulse jets (such as those with thermal or piezoelectric ejectors) are used, with their orifices on a common front surface of the head.
- the method comprises positioning the head with the orifice facing the substrate. Multiple fluid droplets of the biopolymer, biomonomer or other fluid, are dispensed from the head orifice so as to form an array of droplets on the substrate (this formed array may or may not be the same as the final desired array since, for example, multiple heads can be used to form the final array).
- a gas flow is directed through a venturi which has a throat opening communicating with the dispensing head chamber.
- the gas used may be any suitable gas which may be selected depending upon the reagent chemistry. For example, when phosphoramidite oligonucleotide synthesis or other water sensitive chemistries are used, the gas should preferably be an inert anhydrous compressed gas such as anhydrous nitrogen.
- Gas flow rate through the venturi may be adjusted to alter the chamber pressure. This adjustment can occur whenever it is desired to change the pressure in the chamber, for example before or after the dispensing step.
- the adjustment can be accomplished by suitable means such as a valve on the venturi inlet and/or outlet side, or some other way of at least partially obstructing the inlet and/or outlet side (for example, an operator may simply manually block the outlet side). It will be appreciated from this arrangement, that all of the pressures in or at various chambers in the head therefore, are typically gas pressure (that is, provided by a gas in the location specified).
- the venturi throat opening may provide a negative spotting pressure to the head chamber during dispensing of the droplets, and the gas flow resistance of the venturi outlet side may be adjusted (before or after dispensing) to provide a positive chamber pressure.
- This positive pressure may be provided by increasing the gas flow resistance of the venturi outlet side before dispensing (for example, as a priming pressure so as to assist in priming the jets) or after dispensing (for example, as a purging pressure so as to assist in purging any fluid remaining in the chamber out through the orifice).
- the priming and purging pressures may be the same or different, and each will typically be higher than the spotting pressure. In the case of purging, a purge fluid may optionally be added to the head chamber prior to providing the purging pressure.
- the chamber is loaded with the fluid from a direction behind the orifice (that is, liquid is not loaded through the orifice).
- the gas flow resistance of the venturi outlet side is increased to provide a positive priming pressure to the chamber. This assists in forcing liquid into the one or more jets to prime them.
- the method may additionally include, prior to the dispensing step, loading the head with a fluid, such as a fluid containing a biomonomer (for example, a nucleotide reagent), biopolymer (for example, a pre-synthesized oligonucleotide, cDNA, or DNA purified or amplified from a natural source), or other fluid (for example a fluid containing a moiety or a reagent used in producing such chemical a moiety).
- a fluid such as a fluid containing a biomonomer (for example, a nucleotide reagent), biopolymer (for example, a pre-synthesized oligonucleotide, cDNA, or DNA purified or amplified from a natural source), or other fluid (for example a fluid containing a moiety or a reagent used in producing such chemical a moiety).
- a fluid containing a biomonomer for example, a nucleo
- This loading step includes positioning the head facing a load station which is spaced from the substrate, with the one or more orifices adjacent and facing the fluid to be loaded.
- a loading pressure is provided in the chamber from the venturi throat opening while the head is facing the load station, which is sufficiently negative such that the fluid is drawn into the chamber through the one or more orifices.
- the gas flow rate through the venturi is adjusted to provide a spotting pressure to the chamber while dispensing droplets from the head, which spotting pressure may be the same or higher (that is, less negative) than the loading pressure. This adjustment may, for example, be accomplished by adjusting a valve on the inlet side of the venturi.
- the method may include the loading, spotting and purging steps as described above.
- the present invention provides a method of fabricating an array of different moieties, particularly biopolymers on a substrate using a biopolymer or biomonomer fluid, and using a fluid dispensing head as described above, which method includes positioning the head with the orifice facing the substrate. Multiple fluid droplets of the biopolymer, biomonomer, or other fluid are dispensed from the head so as to form an array of droplets on the substrate. A flow of inert anhydrous gas is directed through a venturi which has a throat opening communicating with the dispensing head chamber.
- This aspect may additionally include providing any of the loading, spotting and purging pressures, in the same manners as mentioned above.
- the head used in the method may have multiple pulse jets with orifices on a common front face of the head, such as a typical inkjet printing head.
- some or all of the jets can be loaded with the same or different fluids (biopolymer or otherwise, for example, deprotection reagent or other reagent).
- the apparatus comprises a substrate station on which the substrate can be mounted, and a fluid dispensing head, and venturi, all as described above.
- the apparatus may further include a source of inert anhydrous gas communicating with the venturi pressurized inlet, and/or a valve to adjust the gas flow rate through the venturi (the valve being on the inlet or outlet side of the venturi, or a valve can be provided on both sides).
- a positioning system moves at least one of the dispensing head and mounted substrate with respect to the other, so that multiple droplets dispensed from the head onto the substrate will form an array thereon.
- the apparatus may further include, particularly in the aspect used for deposition of previously obtained biopolymers, the load and purge stations.
- a control processor may be present to operate the positioning system to selectively position the head facing any one of the stations, and which processor also adjusts the venturi outlet control valve to any of the required positions.
- the load station comprises a plate on which multiple drops of different solutions can be retained.
- the method and apparatus of the present invention can provide a simple way of controlling backpressure in a pulse type fluid dispensing head, and can also provide a simple way of purging the head, without requiring an overly complex system of valves.
- the apparatus and method can also facilitate isolating reagents in the head from moisture or other undesirable components.
- FIG. 1 is a perspective view of a substrate bearing multiple arrays, as may be produced by a method and apparatus of the present invention
- FIG. 2 is an enlarged view of a portion of FIG. 1 showing some of the identifiable individual regions of a single array of FIG. 1;
- FIG. 3 is an enlarged cross-section of a portion of FIG. 2;
- FIG. 4 is a schematic view showing components of an apparatus of the present invention.
- FIG. 5 is a schematic view of an apparatus of the present invention utilizing the components of FIG. 4;
- FIG. 6 is another embodiment of an apparatus of the present invention.
- FIG. 7 is an enlarged cross-section of a load station of the apparatus of FIG. 1;
- FIG. 8 is an enlarged cross-section of a purge station of the apparatus of FIG. 1;
- FIG. 9 is an enlarged cross-section of a cleaning station of the apparatus of FIG. 1;
- FIG. 10 is a top view of a fluid dispensing head used in an apparatus of the present invention.
- FIG. 11 is a bottom view of the head of FIG. 7;
- FIG. 12 is a cross-section along the line 9 - 9 in FIG. 7.
- a “biopolymer” includes peptides or polynucleotides, as well as such compounds composed of or containing amino acid or nucleotide analogs or non-nucleotide groups. This includes those compounds in which the conventional polynucleotide backbone has been replaced with a non-naturally occurring or synthetic backbone, and those a nucleic acid in which one or more of the conventional bases has been replaced with a synthetic base capable of participating in Watson-Crick type hydrogen bonding interactions.
- Polynucleotides include single or multiple stranded configurations, where one or more of the strands may or may not be completely aligned with another.
- a “nucleotide” refers to a subunit of a nucleic acid and includes a phosphate group, a 5 carbon sugar and a nitrogen containing base, as well as analogs of such subunits.
- a “biopolymer” includes DNA (including cDNA), RNA and oligonucleotides.
- An “oligonucleotide” generally refers to a nucleotide multimer of about 10 to 100 nucleotides in length, while a “polynucleotide” includes a nucleotide multimer having any number of nucleotides.
- a “biomonomer” references a single unit, which can be linked with the same or other biomonomers to form a biopolymer (for example, a single amino acid or nucleotide with two linking groups one or both of which may have removable protecting groups).
- a biomonomer fluid or biopolymer fluid reference a fluid containing either a biomonomer or biopolymer, respectively.
- An “array”, unless a contrary intention appears, includes any one or two dimensional arrangement of discrete regions bearing particular moieties (for example, different polynucleotide sequences) associated with that region.
- either embodiment of invention described below may produce multiple identical arrays 12 (only some of which are shown in FIG. 1) across the complete surface of a single substrate 14 .
- the arrays 12 produced on a given substrate need not be identical and some or all could be different.
- Each array 12 will contain multiple spots or regions, 16 .
- an array 12 may contain any number of multiple regions, with a typical number being from 100 to 10,000 regions (although more or less are possible). All of the regions 16 may be different, or some or all could be the same. All of the regions 16 may be different, or some or all could be the same.
- Each region carries a predetermined moiety or a predetermined mixture of moieties, such as a particular polynucleotide sequence or a predetermined mixture of polynucleotides. This is illustrated somewhat schematically in FIG. 3 where regions 16 are shown as carrying different polynucleotide sequences.
- the illustrated apparatus includes a fluid dispensing head 209 which is positioned on a reagent reservoir 207 .
- Head 209 is an inkjet type of printing head with multiple jets each having an orifice, a dispensing chamber and an ejector which, when activated, causes a droplet to be ejected from the orifice.
- Head 209 may be of similar construction to head 210 described below in connection with FIGS. 6 and 10- 12 . However, cover 219 and the individual reservoir chambers 222 of head 210 may be omitted.
- Each reservoir 207 may have a capacity of only about 1 or 2 ml.
- a septum 205 allows sealed access to the interior chamber defined by reservoir 207 by means of a syringe.
- Septum 205 may be made of rubber or other suitable resilient material in a known manner.
- a venturi 80 has inlet and outlet sides 82 , 90 , respectively, an outlet opening 91 , and a throat opening which communicates with reservoir 207 and hence communicates with the dispensing chambers of head 209 .
- a flow of an anhydrous inert gas (particularly nitrogen) can be directed through venturi 80 from an adjustable flow regulator 85 (which may be regarded as a type of valve) and compressed nitrogen tank 88 .
- FIG. 5 An apparatus using the components of FIG. 4 is schematically illustrated in FIG. 5. This apparatus is particularly useful for an in situ method of forming polynucleotides on a substrate 10 using a procedure such as mentioned above.
- Four sets of the components shown in FIG. 4 are used in the apparatus of FIG. 5, with the exception that only a single compressed nitrogen tank 88 is used to supply all four adjustable flow regulators 85 .
- a substrate station 20 can receive and retain substrate 10 thereon.
- a flood station 68 is provided to expose an entire substrate with reagents which are used in the in situ process to simultaneously treat all regions 16 during their formation (for example, with deprotection and washing solutions).
- Substrate station 20 is mounted for movement on a carriage 62 in both an “X” and “Y” direction using a suitable transporter (not shown).
- An enclosure 120 is positioned about the dispensing heads 209 and arranged to allow substrate station 20 to move in and out of it. Enclosure 120 permits the head and substrate station to be maintained in a controlled atmosphere environment (such as an anhydrous nitrogen atmosphere provided from a suitable external source). Movement of substrate station 20 and operation of heads 209 and flood station 68 , can all be coordinated by a suitable processor in an analogous manner to that described in connection with the embodiment FIG. 6.
- flow regulators 85 can be manually adjusted to direct an anhydrous nitrogen flow through each venturi 80 from the inlet side 82 to the outlet side 90 .
- the flow rate is sufficient to provide the spotting pressure to each of the four reservoirs 207 .
- Suitable spotting pressures (which will be slightly negative) are mentioned below.
- any water vapor may be present in reservoirs 207 , they can initially be flushed with anhydrous nitrogen by increasing gas flow resistance on the outlet side 90 of each venturi. This can be accomplished such as by briefly manually blocking each outlet opening 91 (an operator can readily use their finger for this purpose since each venturi 80 may only be in the order of less than 10 mm in width or length).
- valve such as valve 94 described in connection with FIG. 6
- outlet side 90 could be used rather than manual blocking.
- This causes a nitrogen flow from regulators 85 to be briefly forced through lines 96 and hence into reservoirs 207 and out the orifices of the heads 209 . Following flushing, the pressure within reservoirs 207 returns to the spotting pressure.
- Each of reservoirs 207 may then be loaded with respective nucleotide reagents through its septum 205 using a syringe. Given the capillary size of the dispensing chambers and orifices of heads 209 , all jets may not be properly primed such that activation of any ejector may not yield a droplet of the expected volume.
- a priming pressure (which is a positive pressure) can be applied to reservoirs 207 . This can be accomplished by again increasing gas flow resistance on the outlet side 90 of venturis 80 , in a manner already described.
- Enclosure 120 is provided with a nitrogen atmosphere and carriage 62 positions substrate 10 within enclosure 120 facing the orifices of heads 209 .
- the processor then controls movement of substrate 10 in the X and Y directions while coordinating activation of the ejectors in heads 209 , such that droplets are dispensed from heads 209 so as to form an initial array or pattern of droplets on substrate 10 .
- a single array or multiple arrays of droplets can be dispensed in this manner with intervening movement of substrate 10 by carriage 62 to flood station 68 for treatment of all deposited spots with reagents, as required.
- the final product is one or more biopolymer arrays on substrate 10 .
- the apparatus includes a substrate station 20 on which can be mounted the substrate 14 .
- Substrate station 20 can include a vacuum chuck connected to a suitable vacuum source (not shown) to retain a mounted substrate 14 without exerting too much pressure thereon, since substrate 14 is often made of glass.
- a load station 30 , purge station 40 , and cleaning station 50 are spaced apart from one another an substrate station 20 .
- Load station 30 can be of any construction with regions which can retain small volumes of different fluids for loading into head 210 .
- it may be a glass surface with different hydrophobic and hydrophilic regions to retain different drops thereon in the hydrophilic regions.
- Purge station 40 has an upper surface defined by a generally rectangular urethane gasket 43 and a region 42 interior of gasket 43 . Interior region 42 communicates with a vacuum line 72 .
- a vacuum source 74 communicates through vacuum line 72 and an electrically controlled valve 70 , which is controlled by a processor 140 through control line 76 .
- Vacuum source may include a suitable vacuum supply (such as a pump) as well as a trap.
- Gasket 43 is dimensioned such that a periphery of a front face of a dispensing head 210 (described in more detail below) can sealingly engage against upper surface 43 with interior region 42 aligned and communicating with the two rows of orifices 214 in head 210 .
- orifices 214 can be placed in communication with vacuum line 72 so that, during a purging step (described further below) vacuum from line 72 can pull fluid out of head 210 through orifices 214 .
- Any processor in the present application including processor 140 , may be a general purpose microprocessor suitably programmed to execute all of the steps required by the present invention, or any hardware or software combination which will perform the required functions.
- Cleaning station 50 can retain an upwardly facing pad 52 which can be saturated with a suitable cleaning fluid.
- a dispensing head 210 (described in more detail below) is retained by a head retainer 208 .
- Head 210 can be positioned to face any one of loading station 30 , substrate station 20 , purge station 40 , or cleaning station 50 by a positioning system.
- the positioning system includes a carriage 62 connected to each of the foregoing stations, a transporter 60 controlled by processor 140 through line 66 , and a second transporter 100 controlled by processor 140 through line 106 .
- Transporter 60 and carriage 62 are used execute one axis positioning of any of the stations 20 , 30 , 40 and 50 facing the dispensing head 210 by moving them in the direction of arrow 63 , while transporter 100 is used to provide two axis adjustment of the position of head 210 in a vertical direction 202 or in the direction 204 . Further, once substrate station 20 has been positioned facing head 210 , transporter 100 will be used to scan head 208 across a mounted substrate 10 , line by line. However, it will be appreciated that both transporters 60 and 100 , or either one of them, with suitable construction, can be used to perform any necessary positioning (including the foregoing scanning) of head 210 with respect to any of the stations.
- Head retainer 208 communicates with a source of purging fluid, such as tank 110 , through line 112 in which is provided an electrically operable valve 114 controlled by processor 140 through control line 116 .
- the apparatus further includes a venturi 80 having an inlet side 82 communicating through line 84 and an adjustable flow regulator 86 , with a source of compressed anhydrous inert gas in the form of nitrogen tank 88 .
- Flow regulator may optionally be adjusted under control of processor 140 through line 87 . Since a flow regulator may be regarded as a type of valve, flow regulator 86 will often be referenced herein as valve 86 .
- An adjustable venturi outlet control valve 94 communicates with a venturi outlet 90 through line 92 .
- Valve 94 is electrically operable by processor 140 through line 98 and may be of any suitable type, such a simple pivoting gate valve.
- a throat opening 89 in venturi 80 communicates with head retainer 208 , and hence head 210 , through line 96 . It will be appreciated that with the foregoing arrangement, selectable negative or positive pressure can be applied to head 210 from throat opening 89 by adjustment of valve 94 only. Optionally, selectable negative pressures can also be applied by adjustment of valve 86 .
- valve 94 can instead (or additionally) be accomplished by adjusting valve 86 , and that corresponding settings of valve 86 (or both valves) for providing the recited negative pressures can be substituted for the negative pressure producing settings of valve 94 .
- valve 94 when valve 94 is at least partially closed to provide a positive pressure in head 210 from throat opening 89 , such pressure will be provided by the anhydrous nitrogen from cylinder 88 thereby avoiding contact of fluid in head 210 with moisture or other contaminants.
- a source of compressed gas other than the anhydrous nitrogen source can be used.
- dispensing station 20 and head 208 and such other components as may be required or desired, can be enclosed in a controlled atmosphere environment (such as a nitrogen fed environment).
- FIGS. 10, 11, show in plan view a particular fluid dispensing head 210 of the apparatus of FIG. 6.
- Head 210 has multiple fluid dispensing jets, and six reservoir chambers 222 and three hundred capillary delivery chambers 217 .
- an orifice member 212 (here an orifice plate) represents a front face of head 210 , and has orifices 214 disposed in two orifice rows 213 , 215 .
- Each orifice 214 can be regarded as part of a delivery chamber 217 , and tapers inwardly away from a delivery chamber 217 toward an open end 214 a of the orifice 214 .
- Prototypes having this configuration were constructed having 150 orifices in each of the orifice rows.
- each fluid pulse jet includes a fluid dispensing chamber 217 , an ejector 224 as described below, and a reservoir chamber 222
- the six reservoir chambers 222 are shared among a number of delivery chambers 217 (that is, each reservoir chamber 222 has multiple delivery chambers 217 ), while each pulse jet, of course, has its own ejector 224 .
- the number of orifices and corresponding ejectors could of course be varied, for example between 10 to 300 or to 500 or more, depending upon their size and materials used to construct head 210 .
- barrier 220 and adhesive 221 Rearward of orifice member 212 are barrier 220 and adhesive 221 , and, resting upon adhesive 221 is reservoir block 218 and resting upon barrier 220 is back member 216 (here a silicon die, as described more fully below), all more readily understood with reference to a rear view as in FIG. 11 and to a sectional view as in FIG. 12.
- the barrier 220 is a photo polymer layer, and accordingly the delivery chambers (for example delivery chambers 217 ) are defined in part by the inner surface 211 of the orifice plate 212 , in part by the front surface of the margin (for example surface 226 ) of the back member 216 , and in part by an edge (for example edge 225 ) of the portion of the photo polymer layer 220 situated between the orifice plate and the back member.
- the delivery chambers for example delivery chambers 217
- the delivery chambers are defined in part by the inner surface 211 of the orifice plate 212 , in part by the front surface of the margin (for example surface 226 ) of the back member 216 , and in part by an edge (for example edge 225 ) of the portion of the photo polymer layer 220 situated between the orifice plate and the back member.
- the reservoirs (for example reservoir 222 ), which are not separate from the delivery chambers, are defined in part by a portion of an edge of the back member (for example edge portion 227 ), and in part by an inner wall (for example wall 228 ) of the reservoir block 218 together with an edge (for example edge 229 ) of the adhesive layer, situated partly between the reservoir block 218 and the orifice plate 212 .
- a cover 219 (removed in FIG. 11; shown in sectional view in FIG. 12) is affixed to the rear surface of the reservoir block 218 , and sealed peripherally (for example by means of an “O” ring 223 ) so that it provides a common enclosure for the reservoirs.
- Cover 219 is provided with a port 221 , permits communication with holder 208 and hence lines 112 and 97 .
- 10 - 12 can be filled with fluid by contacting the exit ends of the orifices with a quantity of the fluid and then lowering the pressure upstream from the orifices by connecting a source of vacuum at the port in the cover, resulting in drawing fluid in an upstream direction through the orifices into the delivery chambers and then into the reservoirs.
- Selected different fluids can be drawn into the different chambers and reservoirs by contacting each orifice group (in fluid communication with a delivery chamber) with a different fluid.
- each orifice 214 on the front surface 226 of the margin of the back member is an ejector 224 (here an electrical resistor operating as a heating element), which is electrically connected to a source of electrical energy which can be controlled to deliver a suitable pulse of electricity to activate the ejector on demand.
- ejector 224 here an electrical resistor operating as a heating element
- the connectors, the source of electrical energy, and the controller are not shown in the Figs.).
- the back member is a silicon die, and the electrical parts (heating element and connectors, for example) are formed using conventional solid state silicon ship manufacturing techniques.
- the various fluid-handling parts of the head 210 generally have the following characteristics.
- the size of each orifice in the orifice plate is one that produces a spot of suitable dimensions on the substrate surface, where the orifice generally has an exit diameter (or exit diagonal depending upon the particular format of the device) in the range about 1 ⁇ m to 1 mm, usually about 5 ⁇ m to 100 ⁇ m, and more usually about 10 ⁇ m to 60 ⁇ m.
- the fluid capacity of the delivery chamber is in the range about 1 pL to 10 nL, usually about 10 pL to 5 nL and more usually about 50 pL to 1.5 nL.
- the front-to-rear thickness of the delivery chamber, defined by the space between the rear surface of the orifice plate and the front surface of the margin of the back plate, may in some embodiments be in the range less than about 100 ⁇ m, for example in prototypes of embodiments shown in the Figures herein, in the range 10 ⁇ m to 60 ⁇ m.
- the heating element will preferably be made out of a material that can deliver a quick energy pulse, and suitable materials include TaAl and the like.
- the thermal element is capable of achieving temperatures sufficient to vaporize a sufficient volume of the fluid in the firing chamber to produce a bubble of suitable dimensions upon actuation of the ejector.
- the heating element is capable of attaining temperatures at least about 100° C., usually at least about 400° C., and more usually at least about 700° C., and the temperature achievable by the heating element may be as high as 1000° C. or higher. It will be appreciated of course, that other ejector types, such as piezoelectric ejectors, could be used instead of a heating element.
- a device as in FIGS. 10 - 12 can be constructed by adaptation of techniques known in the printing art and, particularly, in the art of inkjet device construction. Certain elements of the device of FIGS. 10 - 12 can be adapted from parts of a commercially available thermal inkjet print head device available from Hewlett-Packard Co. as part no. HP51645A. Various other dispensing head designs can be used, such as those described in U.S. patent application entitled “A MULTIPLE RESERVOIR INK JET DEVICE FOR THE FABRICATION OF BIOMOLECULAR ARRAYS” Ser. No. 09/150,507 filed Sep. 9, 1998. That reference and all other references cited in the present application are incorporated herein by reference.
- each orifice 214 and connected capillary delivery chamber 217 are so dimensioned that they can be expected to fill by capillary action when the orifice 214 is brought into contact with the meniscus of a liquid to be loaded into the pulse jet.
- Reservoir chamber 222 is also capillary but it may be non-capillary (by non-capillary is meant that it is so dimensioned that it will not fill by capillary action after delivery chamber 217 has completely filled).
- reservoir chamber 222 is capillary it is distinguishable from the delivery chamber 217 .
- reservoir chamber 222 could be dimensioned such that it is indistinguishable from delivery chamber 217 (in which case the reservoir and delivery chambers may be the same chamber).
- a slightly negative loading pressure can simultaneously be applied to chamber 222 from pressure source 80 during a load step (described further below), which is sufficiently negative such that the fluid is drawn into the reservoir chamber 222 through the delivery chamber 217 while simultaneously being insufficient to result in ambient atmosphere entering the delivery when no further fluid is facing and adjacent the orifice (typically when a drop of liquid to be loaded, has been completely loaded). Otherwise, fluid being loaded into a jet through an orifice would be drawn into delivery chambers 217 with possible loss of prime of the jets.
- the delivery chambers 217 should be completely filled (and preferably along with at least part of reservoir chambers 222 ), with a liquid face or meniscus being maintained within orifice 214 and preferably at the open end 214 a of the orifices 214 . Air entering orifices 214 after loading may result in loss of this condition.
- Venturi 80 in the presence of a sufficient flow of anhydrous nitrogen from tank 88 and valve 86 , is also capable of providing a “spotting pressure” which is slightly negative, but is typically higher (that is, less negative) than the loading pressure, during a dispensing step (described below).
- the spotting pressure is sufficient to retain fluid within the jets in the absence of activation of a given ejector 224 . This can be obtained by processor 140 at least partially closing valve 94 from the load setting to a more restricted spotting position.
- the spotting pressure will typically be a known quantity for a given head 210 or can also be readily determined by experimentation.
- valve 94 can be further closed from the spotting setting to a “cleaning setting” such that throat opening 89 provides in reservoir chamber 222 a holdoff pressure which is sufficiently positive to prevent liquid contacting the orifices 214 during a head cleaning step (described below) from entering delivery chambers 217 through the orifices 214 .
- the holdoff pressure is a gas pressure in the reservoir chamber 217 (that is, there is a gas in the delivery chamber).
- a positive “purging pressure” which is provided to reservoir chamber 222 by providing a negative pressure from pump 74 to purge station 40 , could instead be replaced by providing a positive pressure to reservoir chamber 22 from throat opening 89 by at least partially closing valve 94 .
- the corresponding “purge setting” of valve 94 will typically be a further closed position from the cleaning setting since the purging pressure will typically be higher than the holdoff pressure.
- the loading pressure is a negative pressure which will typically be less than the capillary pressure within a given jet during loading (for example, 10-90% of the capillary pressure), although allowances may need to be made for other factors such as the weight of the fluid column in a jet during loading (although in most fluid heads this will be negligible compared to capillary pressure).
- the mensiscus at an orifice 214 has a capillary pressure based on its curvature. To avoid air (or other ambient gas) from entering a delivery chamber 217 the meniscus should not move away from the end of an orifice 214 . This basically implies that the value of the loading pressure should be below this capillary pressure.
- a suitable loading pressure for any particular apparatus can be readily determined by experimentation, simply by adjusting valve 94 (and/or valve 86 as already mentioned) until the required result is observed. That is, liquid to be loaded is drawn into reservoir chamber 222 without ambient atmosphere outside orifices 214 entering the delivery chamber 217 after the reservoir chambers have been loaded and there is no further fluid facing and adjacent the orifices 214 . When too high a negative pressure is used, entry of ambient atmosphere into delivery chambers 217 can be observed directly or from the fact that the jets have lost their prime. When prime is lost, one way to regain it is to purge the head and reload it.
- the load setting of valve 94 can be recorded by processor 140 or can be set mechanically in valve 94 .
- Suitable spotting, purge and holdoff pressures can also be readily determined by experimentation or calculation, and the corresponding settings of valves 84 , 94 recorded by processor 140 .
- the purging pressure is greater than the holdoff pressure which is greater than the spotting pressure, which is in turn greater than the loading pressure (that is, the spotting pressure is less negative than the loading pressure).
- ambient pressure will typically be about 14.7 psia
- the capillary pressure in a head of the above described type might be about 18 inches of water (0.65 psig)
- the loading pressure might typically be about 8 inches of water below atmosphere (that is, below ambient pressure).
- the holdoff pressure is greater than the capillary pressure, typically about 2 to 3 times the capillary pressure (for example, about 2 psig or 55 inches of water above atmosphere), while the spotting pressure is typically about 10-90% of the capillary pressure (for example, about 5 inches of water, or 0.18 psig, below atmosphere).
- the purging pressure will typically be many times the capillary pressure, for example about 10 to 12 psig or 275-330 inches of water above atmosphere. Description of the pressure adjustments is also provided in U.S. Patent Application entitled“FABRICATING BIOPOLYMER ARRAYS”, by M. Caren et al., assigned to the same assignee as this application, Attorney Docket No. 10990640, filed on the same date as the present application.
- FIGS. 4 through 5 can fabricate arrays of different moieties, including arrays of different biopolymers, such as those illustrated in FIGS. 1 to 3 . Operation of the apparatus to generate biopolymers will now be described although it will be understood that analogous methods can be used to generate arrays of other moieties.
- tank 110 contains a suitable purging fluid (usually a buffered solution), that valve 86 has been manually opened to provide a flow of anhydrous nitrogen so that adjustment of valve 94 (and/or valve 86 , as already mentioned) to the load, spotting, and cleaning settings (and optionally a purge setting) will cause the throat opening to provide the load, spotting and holdoff pressures (and optionally a purging pressure). It will also be assumed that drops of different biomonomer or biopolymer containing fluids (or other fluids) have been placed at respective notches 32 (or other drop retaining regions) of load station 30 .
- a suitable purging fluid usually a buffered solution
- This placement can be accomplished by manual or automated pipetting, or spotting of drops onto load station 30 using glass rods, which are of a volume required to load all of the pulse jets.
- the flexible microtitre plate described in U.S. patent application “Method and Apparatus for Liquid Transfer”, Ser. No. 09/183,604 could be used as load station 30 .
- pad 52 has been previously placed in cleaning station 50 and saturated with a suitable cleaning solution. Operation of the following sequences are controlled by processor 140 unless a contrary indication appears.
- a loading sequence is initiated in which processor 140 directs the positioning system to position head 210 facing load station 30 with the orifices aligned, facing, and adjacent to respective drops on load station 30 .
- processor 140 may optionally ensure valve 86 is open and selects the load setting of valve 94 so that the loading pressure is applied to load chamber 222 and hence to delivery chamber 217 . Capillary pressure will cause fluid to then simultaneously flow in through orifices 214 to fill delivery chambers 217 and reservoir chambers 222 .
- the load pressure assists in this filling by causing the fluid to flow faster. At this point, or shortly thereafter, there will be no further fluid facing and adjacent orifices 214 , either because the fluid at load station 30 is exhausted or head 210 is moved away from load station 30 . In the case where head 210 is moved away before all fluid is exhausted, some fluid may remain on the front face of head 210 and will continue to be drawn in until exhausted. In any event, because of the value of the loading pressure as discussed above, ambient atmosphere (air or nitrogen, for example) does not then enter delivery chambers 217 .
- a dispensing sequence is then initiated in which processor 140 then causes the positioning system to position head 210 facing substrate station 20 , and particularly the mounted substrate 10 , and with head 210 at an appropriate distance from substrate 10 .
- the load setting of valve 94 is selected.
- Processor 140 then activates ejectors 224 in a controlled sequence while causing the positioning system to scan head 210 across substrate 10 line by line (or in some other desired pattern), to dispense droplets in a configuration which results in multiple arrays of the desired configuration on substrate 10 . If necessary or desired, processor 140 can repeat the load and dispensing sequences one or more times.
- a purging sequence is initiated by processor 140 causing the positioning system to position head 210 facing, and in sealing engagement against, purge station such that orifices 214 are in communication with vacuum source 74 .
- Processor 140 selects a neutral position of valves 84 , 94 in which reservoir chamber 222 is essentially open to ambient pressure, and opens valve 114 such that a predetermined quantity of a purge fluid fills chambers 222 , 217 .
- Valve 114 is then closed and valve 70 opened by processor 140 , so that vacuum is thereby applied from outside of orifices 214 resulting in purging of liquid in head 210 simultaneously out through orifices 214 .
- processor 140 causes the positioning system to position head 210 at cleaning station 50 , ensures valve 84 is closed selects the holdoff setting of valve 94 , and causes head 210 to wipe across saturated pad 52 thereby cleaning plate 212 including the regions around the orifices 214 .
- processor 140 causes the positioning system to position head 210 at cleaning station 50 , ensures valve 84 is closed selects the holdoff setting of valve 94 , and causes head 210 to wipe across saturated pad 52 thereby cleaning plate 212 including the regions around the orifices 214 .
- the gas pressure inside delivery chamber 217 exceeds the capillary pressure, some outgassing will occur through orifices 214 (that is, bubbling of gas exiting orifices will be seen there).
- activation results in raising the temperature of the heater to a temperature sufficient to vaporize a small portion of the fluid immediately adjacent the heater and produce a bubble.
- the temperature of the heater is raised to a temperature at least about 100° C., usually at least about 400° and more usually at least about 700° C., and the temperature may be raised as high as 1000° C. or higher, but is usually raised to a temperature that does not exceed about 2000° C. and more usually does not exceed about 1500° C. Accordingly, a sufficient amount of energy will be delivered to the resistive element to produce the requisite temperature rise, where the amount of energy is generally in the range about 1.0 to 100 ⁇ J, usually about 1.5 to 15 ⁇ J.
- the portion of fluid in the firing chamber that is vaporized will be sufficient to produce a bubble in the firing chamber of sufficient volume to force an amount of liquid out of the orifice.
- the formation of the bubble in the firing chamber traps a portion or volume of the fluid present in the firing chamber between the heating element and the orifice and forces an amount or volume of fluid out of the orifice at high speed.
- the amount or volume of fluid that is forced out of the firing chamber can be controlled according to the quantity of biological material to be deposited at the particular location on the receiving surface.
- the amount of fluid that is expelled in a single activation event can be controlled by changing one or more of a number of parameters, including the orifice diameter, the orifice length (thickness of the orifice member at the orifice), the size of the deposition chamber, and the size of the heating element, among others.
- the amount of fluid that is expelled during a single activation event is generally in the range about 0.1 to 1000 pL, usually about 0.5 to 500 pL and more usually about 1.0 to 250 pL.
- a typical velocity at which the fluid is expelled from the chamber is more than about 1 m/s, usually more than about 10 m/s, and may be as great as about 20 m/s or greater.
- the spot dimensions can be controlled such that spots of various sizes can be produced.
- the sizes of the spots can have widths (that is, diameter, for a round spot) in the range from a minimum of about 10 ⁇ m to a maximum of about 1.0 cm.
- material can be deposited according to the invention in small spots whose width is in the range about 1.0 ⁇ m to 1.0 mm, usually about 5.0 ⁇ m to 500 ⁇ m, and more usually about 10 ⁇ m to 200 ⁇ m.
- any of a variety of geometries may be constructed, including for example, organized rows and columns of spots (for example, a grid of spots, across the substrate surface), a series of curvilinear rows across the substrate surface(for example, a series of concentric circles or semi-circles of spots), and the like.
- An array according to the invention generally includes at least tens of features, usually at least hundreds, more usually thousands, and as many as a hundred thousand or more features. Where a pattern of spots of an array is deposited on a substrate surface, the pattern may vary as desired.
- the pattern may be in the form of organized rows and columns of spots (for example, a grid of spots, across the substrate surface), a series of curvilinear rows across the substrate surface(for example, a series of concentric circles or semi-circles of spots), and the like.
- the present methods and apparatus may be used to deposit biopolymers or other moieties on surfaces of any of a variety of different substrates, including both flexible and rigid substrates.
- Preferred materials provide physical support for the deposited material and endure the conditions of the deposition process and of any subsequent treatment or handling or processing that may be encountered in the use of the particular array.
- the array substrate may take any of a variety of configurations ranging from simple to complex. Thus, the substrate could have generally planar form, as for example a slide or plate configuration, such as a rectangular or square or disc.
- the substrate will be shaped generally as a rectangular solid, having a length in the range about 4 mm to 200, usually about 4 mm to 150 mm, more usually about 4 mm to 125 mm; a width in the range about 4 mm to 200 mm, usually about 4 mm to 120 mm and more usually about 4 mm to 80 mm; and a thickness in the range about 0.01 mm to 5.0 mm, usually from about 0.1 mm to 2 mm and more usually from about 0.2 to 1 mm.
- the configuration of the array may be selected according to manufacturing, handling, and use considerations.
- the substrates may be fabricated from any of a variety of materials.
- the materials from which the substrate may be fabricated should ideally exhibit a low level of non-specific binding during hybridization events.
- a material that is transparent to visible and/or UV light it will also be preferable to employ a material that is transparent to visible and/or UV light.
- materials of interest include: nylon, both modified and unmodified, nitrocellulose, polypropylene, and the like, where a nylon membrane, as well as derivatives thereof, may be particularly useful in this embodiment.
- specific materials of interest include: glass; plastics (for example, polytetrafluoroethylene, polypropylene, polystyrene, polycarbonate, and blends thereof, and the like); metals (for example, gold, platinum, and the like).
- the substrate surface onto which the polynucleotide compositions or other moieties is deposited may be smooth or substantially planar, or have irregularities, such as depressions or elevations.
- the surface may be modified with one or more different layers of compounds that serve to modify the properties of the surface in a desirable manner.
- modification layers when present, will generally range in thickness from a monomolecular thickness to about 1 mm, usually from a monomolecular thickness to about 0.1 mm and more usually from a monomolecular thickness to about 0.001 mm.
- Modification layers of interest include: inorganic and organic layers such as metals, metal oxides, polymers, small organic molecules and the like.
- Polymeric layers of interest include layers of: peptides, proteins, polynucleic acids or mimetics thereof (for example, peptide nucleic acids and the like); polysaccharides, phospholipids, polyurethanes, polyesters, polycarbonates, polyureas, polyamides, polyethyleneamines, polyarylene sulfides, polysiloxanes, polyimides, polyacetates, and the like, where the polymers may be hetero- or homopolymeric, and may or may not have separate functional moieties attached thereto (for example, conjugated),
- a venturi 80 provides a simple way of providing negative or positive backpressure in a pulse type fluid dispensing head while conveniently isolating sensitive reagents to contaminants (such as moisture).
- variations and modifications of the above described embodiments of the invention are, of course, possible. Accordingly, the present invention is not limited to such embodiments.
Abstract
A method and apparatus for fabricating an array of biopolymers on a substrate using a biopolymer or biomonomer fluid, and using a fluid dispensing head. The head has at least one jet which can dispense droplets onto a substrate, the jet including a chamber with an orifice, and including an ejector which, when activated, causes a droplet to be ejected from the orifice. The method includes positioning the head with the orifice facing the substrate. Multiple droplets of the biopolymer or biomonomer fluid are dispensed from the head orifice so as to form an array of droplets on the substrate. A gas flow is directed through a venturi which has a throat opening communicating with the dispensing head chamber. A venturi control valve which particularly communicate with an outlet of the venturi, is adjusted to alter the chamber pressure. The venturi may be driven by a source of inert anhydrous compressed gas which assists in maintaining fluid in the head isolated from moisture.
Description
- This invention relates to biopolymer arrays, particularly polynucleotide arrays such as DNA arrays, which are useful in diagnostic, screening, gene expression analysis, and other applications.
- Arrays of biopolymers, such as arrays of peptides or polynucleotides (such as DNA or RNA), are known and are used, for example, as diagnostic or screening tools. Such arrays include regions (sometimes referenced as spots) of usually different sequence biopolymers arranged in a predetermined configuration on a substrate. The arrays, when exposed to a sample, will exhibit a pattern of binding which is indicative of the presence and/or concentration of one or more components of the sample, such as an antigen in the case of a peptide array or a polynucleotide of particular sequence in the case of a polynucleotide array. The binding pattern can be detected, for example, by labeling all potential targets (for example, DNA) in the sample with a suitable label (such as a fluorescent compound), and accurately observing the fluorescence pattern on the array.
- Biopolymer arrays can be fabricated using in situ synthesis methods or deposition of the previously obtained biopolymers. The in situ synthesis methods include those described in U.S. Pat. No. 5,449,754 for synthesizing peptide arrays, as well as
WO 98/41531 and the references cited therein for synthesizing polynucleotides (specifically, DNA). Such in situ synthesis methods can be basically regarded as repeating at each spot the sequence of: (a) deprotecting any previously deposited monomer so that it can now link with a subsequently deposited protected monomer; and (b) depositing a droplet of another protected monomer for linking. Different monomers may be deposited at different regions on the substrate during any one iteration so that the different regions of the completed array will have different desired biopolymer sequences. One or more intermediate further steps may be required in each iteration, such as oxidation, capping and washing steps. The deposition methods basically involve depositing biopolymers at predetermined locations on a substrate which are suitably activated such that the biopolymers can link thereto. Biopolymers of different sequence may be deposited at different regions of the substrate to yield the completed array. Washing or other additional steps may also be used. Reagents used in typical in situ synthesis are water sensitive, and thus the presence of moisture should be eliminated or at least minimized. - Typical procedures known in the art for deposition DNA such as whole oligomers or cDNA, are to load a small volume of DNA in solution on the tip of a pin or in an open capillary and touch the pin or capillary to the surface of the substrate. When the fluid touches the surface, some of the fluid is transferred. The pin or capillary must be washed prior to picking up the next type of DNA for spotting onto the array. This process is repeated for many different sequences and, eventually, the desired array is formed. Alternatively, the DNA can be loaded into an inkjet head and fired onto the substrate. Such a technique has been described, for example, in PCT publications WO 95/25116 and
WO 98/41531, and elsewhere. This method has the advantage of non-contact deposition. Still other methods include pipetting and positive displacement pumps such as the Bio-Dot A/D3000 Dispenser available from Bio-Dot Inc., Irvine, Calif., USA. There are four important design aspects required to fabricate an array of bioplymers such as cDNA's or DNA oligomers. First, the array sensitivity is dependent on having reproducible spots on the substrate. The location of each type of spot must be known and the spotted area should be uniformly coated with the DNA. Second, since DNA is expensive to produce, a minimum amount of the DNA solution should be loaded into any of the transfer mechanisms. Third, any cross contamination of different DNA's must be lower than the sensitivity of the final array as used in a particular assay, to prevent false positive signals. Therefore, the transfer device must be easily cleaned after each type of DNA is deposited or the device must be inexpensive enough to be a disposable. Finally, since the quantity of the assay sample is often limited, it is advantageous to make the spots small and closely spaced. - Similar technologies can be used for in-situ synthesis of biopolymer arrays, such as DNA oligomer arrays, on a solid substrate. In this case, each oligomer is formed nucleotide by nucleotide directly in the desired location on the substrate surface. This process demands repeatable drop size and accurate placement on the substrate. It is advantageous to have an easily cleaned deposition system since some of the reagents have a limited lifetime and must be purged from the system frequently. Since reagents, such as those used in conventional phosphoramidite DNA chemistry may be water sensitive, there is an additional limitation that these chemical reagents do not come in contact with water or water vapor. Therefore, the system must isolate the reagents from any air that may contain water vapor for hours to days during array fabrication. Additionally, the materials selected to construct system must be compatible with the chemical reagents thereby eliminating a lot of organic materials such as rubber.
- Given the above requirements of biopolymer array fabrication, deposition using an inkjet type head is particularly favorable. In particular, inkjet deposition has advantages which include producing very small spot sizes. This allows high-density arrays to be fabricated. Furthermore, the spot size is uniform and reproducible as demonstrated by the successful use of inkjets in printers. Since it is a non-contact technique, ink-jet deposition will not scratch or damage the surface. Ink-jets have very high deposition rate, which facilitates rapid manufacture of arrays.
- However, an ink-jet deposition system used for fabricating a biopolymer array, should meet a number of requirements. Specifically, the inkjet head must be capable of being loaded with very small volumes of DNA solution and function with minimal or no priming of the inkjets. The system should provide for easy purging of the working solution and readily flushed clean when required. When used for in-situ synthesis, the system should be able to to keep reagents isolated from moisture in the surrounding air. Additionally, use of an inkjet head typically requires that a negative backpressure (that is, a pressure behind an orifice of the jet), in the range of one to six inches of water, be supplied to the inkjet head so that the inkjets form repeatable droplets without dripping during times when the jet has not been activated.
- Open-cell foam has been used to provide this negative backpressure in an inkjet printer in a manner disclosed in U.S. Pat. No. 4,771,295, such the capillarity of the foam creates the negative backpressure in an ink reservoir. While this is an easy and economical way to provide the required negative backpressure, the foam cannot be easily purged of the working fluid. A small rubber thimble, similar to an eyedropper, can alternatively be used but the backpressure will vary as the reservoir is depleted. In addition, rubber is incompatible with the chemical reagents typically used in in-situ synthesis. A spring bag reservoir can be designed to control the backpressure of the fluid reservoir, however it requires a large working volume and is therefore not a good choice for the small reservoir volumes required by DNA or other biopolymer array fabrication. A regulated vacuum source could also be used. However, this may permit undesirable components, such as moisture, entering the head particularly during in situ synthesis. Additionally, purging the inkjet head then requires extra valves and a compressed nitrogen (or other suitable gas) source. Gravity is one of the easiest backpressure control means, however the backpressure changes as the fluid height drops and it requires too large a fluid volume to work properly for the small volumes encountered in an inkjet. It would be desirable then, to provide an apparatus and method for fabricating arrays of biopolymers which can use an inkjet type head or other pulse jet head, and which provides for easy purging and cleaning of the head. It would further be desirable that such an apparatus and method provide a simple way of providing a controlled negative backpressure to the head and also provide a simple way of purging the head, without an overly complex system of valves. It would also be desirable that any apparatus and method facilitates isolating reagents in the head from moisture or other undesirable components, and that it is of a compact construction given the small size of other components typically encountered in polynucleotide synthesis.
- The present invention then, provides a method of fabricating an array of different or the same moieties (for example, multiple different chemical compounds) on a substrate using one or more suitable fluids, and using a fluid dispensing head. The invention is particularly useful for the in situ process since it provides the required head pressure while facilitating isolation of reagents from moisture or other undesirable components. However, the invention is also applicable to the direct deposition of polynucleotides. Particularly, the invention provides a method of fabricating an array of biopolymers using a biopolymer containing fluid, or one or more fluids containing a biomonomer. The head has at least one jet which can dispense droplets of a fluid onto a substrate, the jet including a chamber with an orifice, and including an ejector which, when activated, causes a droplet to be ejected from the orifice. The head may particularly be of a type commonly used in inkjet printers, in which a plurality of pulse jets (such as those with thermal or piezoelectric ejectors) are used, with their orifices on a common front surface of the head.
- The method comprises positioning the head with the orifice facing the substrate. Multiple fluid droplets of the biopolymer, biomonomer or other fluid, are dispensed from the head orifice so as to form an array of droplets on the substrate (this formed array may or may not be the same as the final desired array since, for example, multiple heads can be used to form the final array). A gas flow is directed through a venturi which has a throat opening communicating with the dispensing head chamber. The gas used may be any suitable gas which may be selected depending upon the reagent chemistry. For example, when phosphoramidite oligonucleotide synthesis or other water sensitive chemistries are used, the gas should preferably be an inert anhydrous compressed gas such as anhydrous nitrogen. By “inert” in this context is referenced no substantial adverse reaction with a reagent. Gas flow rate through the venturi may be adjusted to alter the chamber pressure. This adjustment can occur whenever it is desired to change the pressure in the chamber, for example before or after the dispensing step. The adjustment can be accomplished by suitable means such as a valve on the venturi inlet and/or outlet side, or some other way of at least partially obstructing the inlet and/or outlet side (for example, an operator may simply manually block the outlet side). It will be appreciated from this arrangement, that all of the pressures in or at various chambers in the head therefore, are typically gas pressure (that is, provided by a gas in the location specified).
- The venturi throat opening may provide a negative spotting pressure to the head chamber during dispensing of the droplets, and the gas flow resistance of the venturi outlet side may be adjusted (before or after dispensing) to provide a positive chamber pressure. This positive pressure may be provided by increasing the gas flow resistance of the venturi outlet side before dispensing (for example, as a priming pressure so as to assist in priming the jets) or after dispensing (for example, as a purging pressure so as to assist in purging any fluid remaining in the chamber out through the orifice). The priming and purging pressures may be the same or different, and each will typically be higher than the spotting pressure. In the case of purging, a purge fluid may optionally be added to the head chamber prior to providing the purging pressure.
- In one aspect of the method, which is particularly useful for (but not limited to) the in situ method, the chamber is loaded with the fluid from a direction behind the orifice (that is, liquid is not loaded through the orifice). Following loading, the gas flow resistance of the venturi outlet side is increased to provide a positive priming pressure to the chamber. This assists in forcing liquid into the one or more jets to prime them.
- In another aspect, which is particularly useful for (but not limited to) the deposition of previously obtained biopolymers, the method may additionally include, prior to the dispensing step, loading the head with a fluid, such as a fluid containing a biomonomer (for example, a nucleotide reagent), biopolymer (for example, a pre-synthesized oligonucleotide, cDNA, or DNA purified or amplified from a natural source), or other fluid (for example a fluid containing a moiety or a reagent used in producing such chemical a moiety). This loading step includes positioning the head facing a load station which is spaced from the substrate, with the one or more orifices adjacent and facing the fluid to be loaded. A loading pressure is provided in the chamber from the venturi throat opening while the head is facing the load station, which is sufficiently negative such that the fluid is drawn into the chamber through the one or more orifices. The gas flow rate through the venturi is adjusted to provide a spotting pressure to the chamber while dispensing droplets from the head, which spotting pressure may be the same or higher (that is, less negative) than the loading pressure. This adjustment may, for example, be accomplished by adjusting a valve on the inlet side of the venturi.
- The method may include the loading, spotting and purging steps as described above.
- In another aspect, the present invention provides a method of fabricating an array of different moieties, particularly biopolymers on a substrate using a biopolymer or biomonomer fluid, and using a fluid dispensing head as described above, which method includes positioning the head with the orifice facing the substrate. Multiple fluid droplets of the biopolymer, biomonomer, or other fluid are dispensed from the head so as to form an array of droplets on the substrate. A flow of inert anhydrous gas is directed through a venturi which has a throat opening communicating with the dispensing head chamber. This aspect may additionally include providing any of the loading, spotting and purging pressures, in the same manners as mentioned above. The head used in the method may have multiple pulse jets with orifices on a common front face of the head, such as a typical inkjet printing head. In this case, some or all of the jets can be loaded with the same or different fluids (biopolymer or otherwise, for example, deprotection reagent or other reagent).
- An apparatus which can be used to execute a method of the present invention is also provided. In one aspect, the apparatus comprises a substrate station on which the substrate can be mounted, and a fluid dispensing head, and venturi, all as described above. The apparatus may further include a source of inert anhydrous gas communicating with the venturi pressurized inlet, and/or a valve to adjust the gas flow rate through the venturi (the valve being on the inlet or outlet side of the venturi, or a valve can be provided on both sides). A positioning system moves at least one of the dispensing head and mounted substrate with respect to the other, so that multiple droplets dispensed from the head onto the substrate will form an array thereon. The apparatus may further include, particularly in the aspect used for deposition of previously obtained biopolymers, the load and purge stations. A control processor may be present to operate the positioning system to selectively position the head facing any one of the stations, and which processor also adjusts the venturi outlet control valve to any of the required positions. In a particular embodiment, the load station comprises a plate on which multiple drops of different solutions can be retained.
- The method and apparatus of the present invention can provide a simple way of controlling backpressure in a pulse type fluid dispensing head, and can also provide a simple way of purging the head, without requiring an overly complex system of valves. The apparatus and method can also facilitate isolating reagents in the head from moisture or other undesirable components.
- FIG. 1 is a perspective view of a substrate bearing multiple arrays, as may be produced by a method and apparatus of the present invention;
- FIG. 2 is an enlarged view of a portion of FIG. 1 showing some of the identifiable individual regions of a single array of FIG. 1;
- FIG. 3 is an enlarged cross-section of a portion of FIG. 2;
- FIG. 4 is a schematic view showing components of an apparatus of the present invention;
- FIG. 5 is a schematic view of an apparatus of the present invention utilizing the components of FIG. 4;
- FIG. 6 is another embodiment of an apparatus of the present invention;
- FIG. 7 is an enlarged cross-section of a load station of the apparatus of FIG. 1;
- FIG. 8 is an enlarged cross-section of a purge station of the apparatus of FIG. 1;
- FIG. 9 is an enlarged cross-section of a cleaning station of the apparatus of FIG. 1;
- FIG. 10 is a top view of a fluid dispensing head used in an apparatus of the present invention;
- FIG. 11 is a bottom view of the head of FIG. 7; and
- FIG. 12 is a cross-section along the line9-9 in FIG. 7.
- To facilitate understanding, identical reference numerals have been used, where practical, to designate corresponding elements that are common to the Figures. Figure components are not drawn to scale.
- In the present application, unless a contrary intention appears, the following terms refer to the indicated characteristics. A “biopolymer” includes peptides or polynucleotides, as well as such compounds composed of or containing amino acid or nucleotide analogs or non-nucleotide groups. This includes those compounds in which the conventional polynucleotide backbone has been replaced with a non-naturally occurring or synthetic backbone, and those a nucleic acid in which one or more of the conventional bases has been replaced with a synthetic base capable of participating in Watson-Crick type hydrogen bonding interactions. Polynucleotides include single or multiple stranded configurations, where one or more of the strands may or may not be completely aligned with another. A “nucleotide” refers to a subunit of a nucleic acid and includes a phosphate group, a 5 carbon sugar and a nitrogen containing base, as well as analogs of such subunits. Specifically, a “biopolymer” includes DNA (including cDNA), RNA and oligonucleotides. An “oligonucleotide” generally refers to a nucleotide multimer of about 10 to 100 nucleotides in length, while a “polynucleotide” includes a nucleotide multimer having any number of nucleotides. A “biomonomer” references a single unit, which can be linked with the same or other biomonomers to form a biopolymer (for example, a single amino acid or nucleotide with two linking groups one or both of which may have removable protecting groups). A biomonomer fluid or biopolymer fluid reference a fluid containing either a biomonomer or biopolymer, respectively. An “array”, unless a contrary intention appears, includes any one or two dimensional arrangement of discrete regions bearing particular moieties (for example, different polynucleotide sequences) associated with that region. It will also be appreciated that throughout the present application, words such as “upper”, “lower” and the like are used with reference to a particular orientation of the apparatus with respect to gravity, but it will be understood that other operating orientations of the apparatus or any of its components, with respect to gravity, are possible. Reference to a “droplet” being dispensed from a pulse jet herein, merely refers to a discrete small quantity of fluid (usually less than about 1000 pL) being dispensed upon a single pulse of the pulse jet (corresponding to a single activation of an ejector) and does not require any particular shape of this discrete quantity. “Fluid” is used herein to reference a liquid. Further, when reference is made in this application to providing a pressure within the dispensing head or a chamber therein this refers, unless a contrary indication appears, to a pressure therein relative to the pressure immediately outside the head pulse jet orifices. Such pressures can be provided by varying the pressure outside the head, or inside the head, or both.
- Referring first to FIG. 1, either embodiment of invention described below may produce multiple identical arrays12 (only some of which are shown in FIG. 1) across the complete surface of a
single substrate 14. However, thearrays 12 produced on a given substrate need not be identical and some or all could be different. Eacharray 12 will contain multiple spots or regions, 16. As mentioned above, anarray 12 may contain any number of multiple regions, with a typical number being from 100 to 10,000 regions (although more or less are possible). All of theregions 16 may be different, or some or all could be the same. All of theregions 16 may be different, or some or all could be the same. Each region carries a predetermined moiety or a predetermined mixture of moieties, such as a particular polynucleotide sequence or a predetermined mixture of polynucleotides. This is illustrated somewhat schematically in FIG. 3 whereregions 16 are shown as carrying different polynucleotide sequences. - Referring to FIG. 4 the illustrated apparatus includes a
fluid dispensing head 209 which is positioned on areagent reservoir 207.Head 209 is an inkjet type of printing head with multiple jets each having an orifice, a dispensing chamber and an ejector which, when activated, causes a droplet to be ejected from the orifice.Head 209 may be of similar construction to head 210 described below in connection with FIGS. 6 and 10-12. However, cover 219 and theindividual reservoir chambers 222 ofhead 210 may be omitted. Eachreservoir 207 may have a capacity of only about 1 or 2 ml. Aseptum 205 allows sealed access to the interior chamber defined byreservoir 207 by means of a syringe.Septum 205 may be made of rubber or other suitable resilient material in a known manner. Aventuri 80 has inlet and outlet sides 82, 90, respectively, anoutlet opening 91, and a throat opening which communicates withreservoir 207 and hence communicates with the dispensing chambers ofhead 209. A flow of an anhydrous inert gas (particularly nitrogen) can be directed throughventuri 80 from an adjustable flow regulator 85 (which may be regarded as a type of valve) andcompressed nitrogen tank 88. - An apparatus using the components of FIG. 4 is schematically illustrated in FIG. 5. This apparatus is particularly useful for an in situ method of forming polynucleotides on a
substrate 10 using a procedure such as mentioned above. Four sets of the components shown in FIG. 4 are used in the apparatus of FIG. 5, with the exception that only a singlecompressed nitrogen tank 88 is used to supply all four adjustable flow regulators 85. Asubstrate station 20 can receive and retainsubstrate 10 thereon. Aflood station 68 is provided to expose an entire substrate with reagents which are used in the in situ process to simultaneously treat allregions 16 during their formation (for example, with deprotection and washing solutions).Substrate station 20 is mounted for movement on acarriage 62 in both an “X” and “Y” direction using a suitable transporter (not shown). Anenclosure 120 is positioned about the dispensing heads 209 and arranged to allowsubstrate station 20 to move in and out of it.Enclosure 120 permits the head and substrate station to be maintained in a controlled atmosphere environment (such as an anhydrous nitrogen atmosphere provided from a suitable external source). Movement ofsubstrate station 20 and operation ofheads 209 andflood station 68, can all be coordinated by a suitable processor in an analogous manner to that described in connection with the embodiment FIG. 6. - In operation of the apparatus of FIG. 5, flow
regulators 85 can be manually adjusted to direct an anhydrous nitrogen flow through each venturi 80 from theinlet side 82 to theoutlet side 90. The flow rate is sufficient to provide the spotting pressure to each of the fourreservoirs 207. Suitable spotting pressures (which will be slightly negative) are mentioned below. In the event that any water vapor may be present inreservoirs 207, they can initially be flushed with anhydrous nitrogen by increasing gas flow resistance on theoutlet side 90 of each venturi. This can be accomplished such as by briefly manually blocking each outlet opening 91 (an operator can readily use their finger for this purpose since eachventuri 80 may only be in the order of less than 10 mm in width or length). Alternatively, a valve (such asvalve 94 described in connection with FIG. 6) communicating withoutlet side 90, could be used rather than manual blocking. This causes a nitrogen flow fromregulators 85 to be briefly forced throughlines 96 and hence intoreservoirs 207 and out the orifices of theheads 209. Following flushing, the pressure withinreservoirs 207 returns to the spotting pressure. Each ofreservoirs 207 may then be loaded with respective nucleotide reagents through itsseptum 205 using a syringe. Given the capillary size of the dispensing chambers and orifices ofheads 209, all jets may not be properly primed such that activation of any ejector may not yield a droplet of the expected volume. To ensure proper priming, a priming pressure (which is a positive pressure) can be applied toreservoirs 207. This can be accomplished by again increasing gas flow resistance on theoutlet side 90 ofventuris 80, in a manner already described.Enclosure 120 is provided with a nitrogen atmosphere andcarriage 62positions substrate 10 withinenclosure 120 facing the orifices ofheads 209. The processor then controls movement ofsubstrate 10 in the X and Y directions while coordinating activation of the ejectors inheads 209, such that droplets are dispensed fromheads 209 so as to form an initial array or pattern of droplets onsubstrate 10. A single array or multiple arrays of droplets can be dispensed in this manner with intervening movement ofsubstrate 10 bycarriage 62 toflood station 68 for treatment of all deposited spots with reagents, as required. The final product is one or more biopolymer arrays onsubstrate 10. - Typically, there is sufficient capacity within
reservoirs 207 such thatmany substrates 10 can be treated in turn by the above method to form many arrays. At some point though, it becomes necessary to flushreservoirs 207 and heads 209. Gas flow resistance on theoutlet side 90 of each venturi can then be increased in a manner already described. As already mentioned, this will cause nitrogen to flow throughlines 96 and increase pressure withinreservoirs 207 to what is referenced as a “purging pressure”, so as to force any liquid withinreservoirs 207 out through the orifices inheads 209. This purging pressure is, in practice, a positive pressure which may be about the same as the priming pressure.Openings 91 can be uncovered when all liquid appears to have been purged fromreservoirs 207. A syringe can then be used to add a purge fluid (such as a solvent) to eachreservoir 207 throughsepta 205, and the foregoing purging procedure repeated. - Referring now to the embodiment of FIG. 6 the apparatus includes a
substrate station 20 on which can be mounted thesubstrate 14.Substrate station 20 can include a vacuum chuck connected to a suitable vacuum source (not shown) to retain a mountedsubstrate 14 without exerting too much pressure thereon, sincesubstrate 14 is often made of glass. Aload station 30,purge station 40, and cleaningstation 50 are spaced apart from one another ansubstrate station 20.Load station 30 can be of any construction with regions which can retain small volumes of different fluids for loading intohead 210. For example, it may be a glass surface with different hydrophobic and hydrophilic regions to retain different drops thereon in the hydrophilic regions. Alternatively, the flexible microtitre plate described in U.S. patent application “Method and Apparatus for Liquid Transfer”, Ser. No. 09/183,604 could be used. In thedrawings load station 30 and has an upper surface withsmall notches 32 to assist in retaining multiple individual drops of a biomonomer or biopolymer fluid on that surface. The number ofnotches 32 or other regions for retaining drops of different fluids, is at least equal to (and can be greater than) the number ofreservoir chambers 222 in a printer head 210 (described further below), and are spaced to align withorifices 214 inhead 210. Even where the number of such fluid retaining regions is less than the number oforifices 214, all delivery chambers communicating with one another (through a reservoir chamber 222) can still be filled in the present invention. This occurs since, with the previously defined load pressure value, fluid which has entered areservoir chamber 222 through oneorifice 214 can still be drawn by capillary pressure into other delivery chambers communicating with thesame reservoir chamber 222. -
Purge station 40 has an upper surface defined by a generallyrectangular urethane gasket 43 and aregion 42 interior ofgasket 43.Interior region 42 communicates with avacuum line 72. Avacuum source 74 communicates throughvacuum line 72 and an electrically controlledvalve 70, which is controlled by aprocessor 140 throughcontrol line 76. Vacuum source may include a suitable vacuum supply (such as a pump) as well as a trap.Gasket 43 is dimensioned such that a periphery of a front face of a dispensing head 210 (described in more detail below) can sealingly engage againstupper surface 43 withinterior region 42 aligned and communicating with the two rows oforifices 214 inhead 210. In this manner,orifices 214 can be placed in communication withvacuum line 72 so that, during a purging step (described further below) vacuum fromline 72 can pull fluid out ofhead 210 throughorifices 214. Any processor in the present application, includingprocessor 140, may be a general purpose microprocessor suitably programmed to execute all of the steps required by the present invention, or any hardware or software combination which will perform the required functions. -
Cleaning station 50 can retain an upwardly facingpad 52 which can be saturated with a suitable cleaning fluid. A dispensing head 210 (described in more detail below) is retained by ahead retainer 208.Head 210 can be positioned to face any one ofloading station 30,substrate station 20,purge station 40, or cleaningstation 50 by a positioning system. The positioning system includes acarriage 62 connected to each of the foregoing stations, atransporter 60 controlled byprocessor 140 throughline 66, and asecond transporter 100 controlled byprocessor 140 throughline 106.Transporter 60 andcarriage 62 are used execute one axis positioning of any of thestations head 210 by moving them in the direction ofarrow 63, whiletransporter 100 is used to provide two axis adjustment of the position ofhead 210 in avertical direction 202 or in thedirection 204. Further, oncesubstrate station 20 has been positioned facinghead 210,transporter 100 will be used to scanhead 208 across a mountedsubstrate 10, line by line. However, it will be appreciated that bothtransporters head 210 with respect to any of the stations. Thus, when the present application recites “positioning” one element (such as head 210) in relation to another element (such as one of thestations -
Head retainer 208, and hence head 210 (specifically,delivery chambers 217 ofhead 210 as described below), communicates with a source of purging fluid, such astank 110, throughline 112 in which is provided an electricallyoperable valve 114 controlled byprocessor 140 throughcontrol line 116. The apparatus further includes aventuri 80 having aninlet side 82 communicating throughline 84 and anadjustable flow regulator 86, with a source of compressed anhydrous inert gas in the form ofnitrogen tank 88. Flow regulator may optionally be adjusted under control ofprocessor 140 throughline 87. Since a flow regulator may be regarded as a type of valve,flow regulator 86 will often be referenced herein asvalve 86. An adjustable venturioutlet control valve 94 communicates with aventuri outlet 90 throughline 92.Valve 94 is electrically operable byprocessor 140 throughline 98 and may be of any suitable type, such a simple pivoting gate valve. Athroat opening 89 inventuri 80 communicates withhead retainer 208, and hence head 210, throughline 96. It will be appreciated that with the foregoing arrangement, selectable negative or positive pressure can be applied to head 210 from throat opening 89 by adjustment ofvalve 94 only. Optionally, selectable negative pressures can also be applied by adjustment ofvalve 86. Thus, in the discussion below where negative pressures are varied by adjustingvalve 94, it will be understood that this can instead (or additionally) be accomplished by adjustingvalve 86, and that corresponding settings of valve 86 (or both valves) for providing the recited negative pressures can be substituted for the negative pressure producing settings ofvalve 94. Furthermore, it will be appreciated that whenvalve 94 is at least partially closed to provide a positive pressure inhead 210 from throat opening 89, such pressure will be provided by the anhydrous nitrogen fromcylinder 88 thereby avoiding contact of fluid inhead 210 with moisture or other contaminants. However, if the fluids in head are not sensitive to moisture or other particular gasses, a source of compressed gas other than the anhydrous nitrogen source can be used. When the fluids are moisture or otherwise sensitive, dispensingstation 20 andhead 208, and such other components as may be required or desired, can be enclosed in a controlled atmosphere environment (such as a nitrogen fed environment). - Referring now to Figures FIGS. 10, 11, these show in plan view a particular
fluid dispensing head 210 of the apparatus of FIG. 6.Head 210 has multiple fluid dispensing jets, and sixreservoir chambers 222 and three hundredcapillary delivery chambers 217. In a front view, FIG. 10, an orifice member 212 (here an orifice plate) represents a front face ofhead 210, and hasorifices 214 disposed in twoorifice rows orifice 214 can be regarded as part of adelivery chamber 217, and tapers inwardly away from adelivery chamber 217 toward anopen end 214 a of theorifice 214. Prototypes having this configuration were constructed having 150 orifices in each of the orifice rows. Thus, while each fluid pulse jet includes afluid dispensing chamber 217, anejector 224 as described below, and areservoir chamber 222, the sixreservoir chambers 222 are shared among a number of delivery chambers 217 (that is, eachreservoir chamber 222 has multiple delivery chambers 217), while each pulse jet, of course, has itsown ejector 224. It will be appreciated that the number of orifices and corresponding ejectors could of course be varied, for example between 10 to 300 or to 500 or more, depending upon their size and materials used to constructhead 210. Rearward oforifice member 212 arebarrier 220 and adhesive 221, and, resting upon adhesive 221 isreservoir block 218 and resting uponbarrier 220 is back member 216 (here a silicon die, as described more fully below), all more readily understood with reference to a rear view as in FIG. 11 and to a sectional view as in FIG. 12. - In a particular configuration, the
barrier 220 is a photo polymer layer, and accordingly the delivery chambers (for example delivery chambers 217) are defined in part by theinner surface 211 of theorifice plate 212, in part by the front surface of the margin (for example surface 226) of theback member 216, and in part by an edge (for example edge 225) of the portion of thephoto polymer layer 220 situated between the orifice plate and the back member. And, in such a configuration, the reservoirs (for example reservoir 222), which are not separate from the delivery chambers, are defined in part by a portion of an edge of the back member (for example edge portion 227), and in part by an inner wall (for example wall 228) of thereservoir block 218 together with an edge (for example edge 229) of the adhesive layer, situated partly between thereservoir block 218 and theorifice plate 212. - A cover219 (removed in FIG. 11; shown in sectional view in FIG. 12) is affixed to the rear surface of the
reservoir block 218, and sealed peripherally (for example by means of an “O” ring 223) so that it provides a common enclosure for the reservoirs. Cover 219 is provided with aport 221, permits communication withholder 208 and hencelines 112 and 97. As will be appreciated, the delivery chambers and reservoirs of the device of FIGS. 10-12 can be filled with fluid by contacting the exit ends of the orifices with a quantity of the fluid and then lowering the pressure upstream from the orifices by connecting a source of vacuum at the port in the cover, resulting in drawing fluid in an upstream direction through the orifices into the delivery chambers and then into the reservoirs. Selected different fluids (or fluids containing different materials) can be drawn into the different chambers and reservoirs by contacting each orifice group (in fluid communication with a delivery chamber) with a different fluid. - Opposite each
orifice 214 on thefront surface 226 of the margin of the back member is an ejector 224 (here an electrical resistor operating as a heating element), which is electrically connected to a source of electrical energy which can be controlled to deliver a suitable pulse of electricity to activate the ejector on demand. (The connectors, the source of electrical energy, and the controller are not shown in the Figs.). In a particular embodiment the back member is a silicon die, and the electrical parts (heating element and connectors, for example) are formed using conventional solid state silicon ship manufacturing techniques. - The various fluid-handling parts of the
head 210 generally have the following characteristics. The size of each orifice in the orifice plate is one that produces a spot of suitable dimensions on the substrate surface, where the orifice generally has an exit diameter (or exit diagonal depending upon the particular format of the device) in the range about 1 μm to 1 mm, usually about 5 μm to 100 μm, and more usually about 10 μm to 60 μm. The fluid capacity of the delivery chamber is in the range about 1 pL to 10 nL, usually about 10 pL to 5 nL and more usually about 50 pL to 1.5 nL. Thereservoir chamber 222 and the connecteddelivery chamber 217, with which any one of theorifices 214 communicate, together have a combined fluid capacity in the range about 1 pL up to 1 mL (more typically less than 100 μL), usually about 0.5 μL to 10 μL, and more usually about 1 μL to 5 μL. The front-to-rear thickness of the delivery chamber, defined by the space between the rear surface of the orifice plate and the front surface of the margin of the back plate, may in some embodiments be in the range less than about 100 μm, for example in prototypes of embodiments shown in the Figures herein, in therange 10 μm to 60 μm. - Where the ejector is a heating element, the heating element will preferably be made out of a material that can deliver a quick energy pulse, and suitable materials include TaAl and the like. The thermal element is capable of achieving temperatures sufficient to vaporize a sufficient volume of the fluid in the firing chamber to produce a bubble of suitable dimensions upon actuation of the ejector. Generally, the heating element is capable of attaining temperatures at least about 100° C., usually at least about 400° C., and more usually at least about 700° C., and the temperature achievable by the heating element may be as high as 1000° C. or higher. It will be appreciated of course, that other ejector types, such as piezoelectric ejectors, could be used instead of a heating element.
- A device as in FIGS.10-12 can be constructed by adaptation of techniques known in the printing art and, particularly, in the art of inkjet device construction. Certain elements of the device of FIGS. 10-12 can be adapted from parts of a commercially available thermal inkjet print head device available from Hewlett-Packard Co. as part no. HP51645A. Various other dispensing head designs can be used, such as those described in U.S. patent application entitled “A MULTIPLE RESERVOIR INK JET DEVICE FOR THE FABRICATION OF BIOMOLECULAR ARRAYS” Ser. No. 09/150,507 filed Sep. 9, 1998. That reference and all other references cited in the present application are incorporated herein by reference.
- It should be noted that the above dimensions of the
head 210, and particularly the dimensions of the deliver chamber 217 (and included orifices 214) are small enough that capillary forces can have a significant effect on the fluid pressures within the fluid column contained within these and larger fluid-handling parts. Particularly, eachorifice 214 and connectedcapillary delivery chamber 217 are so dimensioned that they can be expected to fill by capillary action when theorifice 214 is brought into contact with the meniscus of a liquid to be loaded into the pulse jet.Reservoir chamber 222 is also capillary but it may be non-capillary (by non-capillary is meant that it is so dimensioned that it will not fill by capillary action afterdelivery chamber 217 has completely filled). Whilereservoir chamber 222 is capillary it is distinguishable from thedelivery chamber 217. However,reservoir chamber 222 could be dimensioned such that it is indistinguishable from delivery chamber 217 (in which case the reservoir and delivery chambers may be the same chamber). A slightly negative loading pressure can simultaneously be applied tochamber 222 frompressure source 80 during a load step (described further below), which is sufficiently negative such that the fluid is drawn into thereservoir chamber 222 through thedelivery chamber 217 while simultaneously being insufficient to result in ambient atmosphere entering the delivery when no further fluid is facing and adjacent the orifice (typically when a drop of liquid to be loaded, has been completely loaded). Otherwise, fluid being loaded into a jet through an orifice would be drawn intodelivery chambers 217 with possible loss of prime of the jets. That is, thedelivery chambers 217 should be completely filled (and preferably along with at least part of reservoir chambers 222), with a liquid face or meniscus being maintained withinorifice 214 and preferably at theopen end 214 a of theorifices 214.Air entering orifices 214 after loading may result in loss of this condition. -
Venturi 80, in the presence of a sufficient flow of anhydrous nitrogen fromtank 88 andvalve 86, is also capable of providing a “spotting pressure” which is slightly negative, but is typically higher (that is, less negative) than the loading pressure, during a dispensing step (described below). The spotting pressure is sufficient to retain fluid within the jets in the absence of activation of a givenejector 224. This can be obtained byprocessor 140 at least partially closingvalve 94 from the load setting to a more restricted spotting position. The spotting pressure will typically be a known quantity for a givenhead 210 or can also be readily determined by experimentation. Additionally,valve 94 can be further closed from the spotting setting to a “cleaning setting” such thatthroat opening 89 provides in reservoir chamber 222 a holdoff pressure which is sufficiently positive to prevent liquid contacting theorifices 214 during a head cleaning step (described below) from enteringdelivery chambers 217 through theorifices 214. The holdoff pressure is a gas pressure in the reservoir chamber 217 (that is, there is a gas in the delivery chamber). It will also be appreciated that during a purge step for head 210 (described below), a positive “purging pressure” which is provided toreservoir chamber 222 by providing a negative pressure frompump 74 to purgestation 40, could instead be replaced by providing a positive pressure toreservoir chamber 22 from throat opening 89 by at least partially closingvalve 94. In this case the corresponding “purge setting” ofvalve 94 will typically be a further closed position from the cleaning setting since the purging pressure will typically be higher than the holdoff pressure. - The loading pressure is a negative pressure which will typically be less than the capillary pressure within a given jet during loading (for example, 10-90% of the capillary pressure), although allowances may need to be made for other factors such as the weight of the fluid column in a jet during loading (although in most fluid heads this will be negligible compared to capillary pressure). The mensiscus at an
orifice 214 has a capillary pressure based on its curvature. To avoid air (or other ambient gas) from entering adelivery chamber 217 the meniscus should not move away from the end of anorifice 214. This basically implies that the value of the loading pressure should be below this capillary pressure. A suitable loading pressure for any particular apparatus can be readily determined by experimentation, simply by adjusting valve 94 (and/orvalve 86 as already mentioned) until the required result is observed. That is, liquid to be loaded is drawn intoreservoir chamber 222 without ambient atmosphere outsideorifices 214 entering thedelivery chamber 217 after the reservoir chambers have been loaded and there is no further fluid facing and adjacent theorifices 214. When too high a negative pressure is used, entry of ambient atmosphere intodelivery chambers 217 can be observed directly or from the fact that the jets have lost their prime. When prime is lost, one way to regain it is to purge the head and reload it. The load setting ofvalve 94 can be recorded byprocessor 140 or can be set mechanically invalve 94. Suitable spotting, purge and holdoff pressures can also be readily determined by experimentation or calculation, and the corresponding settings ofvalves processor 140. Generally, the purging pressure is greater than the holdoff pressure which is greater than the spotting pressure, which is in turn greater than the loading pressure (that is, the spotting pressure is less negative than the loading pressure). For example, ambient pressure will typically be about 14.7 psia, the capillary pressure in a head of the above described type might be about 18 inches of water (0.65 psig), while the loading pressure might typically be about 8 inches of water below atmosphere (that is, below ambient pressure). The holdoff pressure is greater than the capillary pressure, typically about 2 to 3 times the capillary pressure (for example, about 2 psig or 55 inches of water above atmosphere), while the spotting pressure is typically about 10-90% of the capillary pressure (for example, about 5 inches of water, or 0.18 psig, below atmosphere). The purging pressure will typically be many times the capillary pressure, for example about 10 to 12 psig or 275-330 inches of water above atmosphere. Description of the pressure adjustments is also provided in U.S. Patent Application entitled“FABRICATING BIOPOLYMER ARRAYS”, by M. Caren et al., assigned to the same assignee as this application, Attorney Docket No. 10990640, filed on the same date as the present application. - The apparatus of FIGS. 4 through 5 can fabricate arrays of different moieties, including arrays of different biopolymers, such as those illustrated in FIGS.1 to 3. Operation of the apparatus to generate biopolymers will now be described although it will be understood that analogous methods can be used to generate arrays of other moieties. First, it will be assumed that
tank 110 contains a suitable purging fluid (usually a buffered solution), thatvalve 86 has been manually opened to provide a flow of anhydrous nitrogen so that adjustment of valve 94 (and/orvalve 86, as already mentioned) to the load, spotting, and cleaning settings (and optionally a purge setting) will cause the throat opening to provide the load, spotting and holdoff pressures (and optionally a purging pressure). It will also be assumed that drops of different biomonomer or biopolymer containing fluids (or other fluids) have been placed at respective notches 32 (or other drop retaining regions) ofload station 30. This placement can be accomplished by manual or automated pipetting, or spotting of drops ontoload station 30 using glass rods, which are of a volume required to load all of the pulse jets. Alternatively, as already mentioned, the flexible microtitre plate described in U.S. patent application “Method and Apparatus for Liquid Transfer”, Ser. No. 09/183,604 could be used asload station 30. Also,pad 52 has been previously placed in cleaningstation 50 and saturated with a suitable cleaning solution. Operation of the following sequences are controlled byprocessor 140 unless a contrary indication appears. - A loading sequence is initiated in which
processor 140 directs the positioning system to positionhead 210 facingload station 30 with the orifices aligned, facing, and adjacent to respective drops onload station 30. As previously mentioned, during any positioning operation one axis positioning of thehead 210 facing the required station can be accomplished throughtransporter 60, and then the other two axes positioning ofhead 210 can be accomplished throughtransporter 100.Processor 140 may optionally ensurevalve 86 is open and selects the load setting ofvalve 94 so that the loading pressure is applied to loadchamber 222 and hence todelivery chamber 217. Capillary pressure will cause fluid to then simultaneously flow in throughorifices 214 to filldelivery chambers 217 andreservoir chambers 222. The load pressure assists in this filling by causing the fluid to flow faster. At this point, or shortly thereafter, there will be no further fluid facing andadjacent orifices 214, either because the fluid atload station 30 is exhausted orhead 210 is moved away fromload station 30. In the case wherehead 210 is moved away before all fluid is exhausted, some fluid may remain on the front face ofhead 210 and will continue to be drawn in until exhausted. In any event, because of the value of the loading pressure as discussed above, ambient atmosphere (air or nitrogen, for example) does not then enterdelivery chambers 217. - A dispensing sequence is then initiated in which
processor 140 then causes the positioning system to positionhead 210 facingsubstrate station 20, and particularly the mountedsubstrate 10, and withhead 210 at an appropriate distance fromsubstrate 10. The load setting ofvalve 94 is selected.Processor 140 then activatesejectors 224 in a controlled sequence while causing the positioning system to scanhead 210 acrosssubstrate 10 line by line (or in some other desired pattern), to dispense droplets in a configuration which results in multiple arrays of the desired configuration onsubstrate 10. If necessary or desired,processor 140 can repeat the load and dispensing sequences one or more times. - Following a dispensing sequence, a purging sequence is initiated by
processor 140 causing the positioning system to positionhead 210 facing, and in sealing engagement against, purge station such thatorifices 214 are in communication withvacuum source 74.Processor 140 selects a neutral position ofvalves reservoir chamber 222 is essentially open to ambient pressure, and opensvalve 114 such that a predetermined quantity of a purge fluid fillschambers Valve 114 is then closed andvalve 70 opened byprocessor 140, so that vacuum is thereby applied from outside oforifices 214 resulting in purging of liquid inhead 210 simultaneously out throughorifices 214. After a suitable predetermined time has elapsed to allow complete purging ofhead 210,processor 140 causes the positioning system to positionhead 210 at cleaningstation 50, ensuresvalve 84 is closed selects the holdoff setting ofvalve 94, and causeshead 210 to wipe across saturatedpad 52 thereby cleaningplate 212 including the regions around theorifices 214. During such operation, since the gas pressure insidedelivery chamber 217 exceeds the capillary pressure, some outgassing will occur through orifices 214 (that is, bubbling of gas exiting orifices will be seen there). - The above sequences can be repeated as often as desired for a
single substrate 10 or multiple different substrates (which may be manually or automatically mounted and held on substrate station 20). - Where the ejectors are electrically resistive heating elements, activation results in raising the temperature of the heater to a temperature sufficient to vaporize a small portion of the fluid immediately adjacent the heater and produce a bubble. The temperature of the heater is raised to a temperature at least about 100° C., usually at least about 400° and more usually at least about 700° C., and the temperature may be raised as high as 1000° C. or higher, but is usually raised to a temperature that does not exceed about 2000° C. and more usually does not exceed about 1500° C. Accordingly, a sufficient amount of energy will be delivered to the resistive element to produce the requisite temperature rise, where the amount of energy is generally in the range about 1.0 to 100 μJ, usually about 1.5 to 15 μJ. The portion of fluid in the firing chamber that is vaporized will be sufficient to produce a bubble in the firing chamber of sufficient volume to force an amount of liquid out of the orifice.
- The formation of the bubble in the firing chamber traps a portion or volume of the fluid present in the firing chamber between the heating element and the orifice and forces an amount or volume of fluid out of the orifice at high speed. The amount or volume of fluid that is forced out of the firing chamber can be controlled according to the quantity of biological material to be deposited at the particular location on the receiving surface. As is well known in the ink jet print art, the amount of fluid that is expelled in a single activation event can be controlled by changing one or more of a number of parameters, including the orifice diameter, the orifice length (thickness of the orifice member at the orifice), the size of the deposition chamber, and the size of the heating element, among others. The amount of fluid that is expelled during a single activation event is generally in the range about 0.1 to 1000 pL, usually about 0.5 to 500 pL and more usually about 1.0 to 250 pL. A typical velocity at which the fluid is expelled from the chamber is more than about 1 m/s, usually more than about 10 m/s, and may be as great as about 20 m/s or greater. As will be appreciated, if the orifice is in motion with respect to the receiving surface at the time an ejector is activated, the actual site of deposition of the material will not be the location that is at the moment of activation in a line-of-sight relation to the orifice, but will be a location that is predictable for the given distances and velocities.
- Upon actuation of an ejector, as described above, fluid is expelled from the orifice and travels to the substrate surface, where it forms a spot on the substrate surface. In this manner, the biological material (such as a nucleic acid) is deposited on the substrate surface. As mentioned above, by varying the operating parameters of the apparatus, the spot dimensions can be controlled such that spots of various sizes can be produced. The sizes of the spots (and, hence, of the array features) can have widths (that is, diameter, for a round spot) in the range from a minimum of about 10 μm to a maximum of about 1.0 cm. In embodiments where very small spot sizes or feature sizes are desired, material can be deposited according to the invention in small spots whose width is in the range about 1.0 μm to 1.0 mm, usually about 5.0 μm to 500 μm, and more usually about 10 μm to 200 μm.
- Where a pattern of arrays is desired, any of a variety of geometries may be constructed, including for example, organized rows and columns of spots (for example, a grid of spots, across the substrate surface), a series of curvilinear rows across the substrate surface(for example, a series of concentric circles or semi-circles of spots), and the like. An array according to the invention generally includes at least tens of features, usually at least hundreds, more usually thousands, and as many as a hundred thousand or more features. Where a pattern of spots of an array is deposited on a substrate surface, the pattern may vary as desired. As such, the pattern may be in the form of organized rows and columns of spots (for example, a grid of spots, across the substrate surface), a series of curvilinear rows across the substrate surface(for example, a series of concentric circles or semi-circles of spots), and the like.
- The present methods and apparatus may be used to deposit biopolymers or other moieties on surfaces of any of a variety of different substrates, including both flexible and rigid substrates. Preferred materials provide physical support for the deposited material and endure the conditions of the deposition process and of any subsequent treatment or handling or processing that may be encountered in the use of the particular array. The array substrate may take any of a variety of configurations ranging from simple to complex. Thus, the substrate could have generally planar form, as for example a slide or plate configuration, such as a rectangular or square or disc. In many embodiments, the substrate will be shaped generally as a rectangular solid, having a length in the range about 4 mm to 200, usually about 4 mm to 150 mm, more usually about 4 mm to 125 mm; a width in the range about 4 mm to 200 mm, usually about 4 mm to 120 mm and more usually about 4 mm to 80 mm; and a thickness in the range about 0.01 mm to 5.0 mm, usually from about 0.1 mm to 2 mm and more usually from about 0.2 to 1 mm. The configuration of the array may be selected according to manufacturing, handling, and use considerations.
- The substrates may be fabricated from any of a variety of materials. In certain embodiments, such as for example where production of binding pair arrays for use in research and related applications is desired, the materials from which the substrate may be fabricated should ideally exhibit a low level of non-specific binding during hybridization events. In many situations, it will also be preferable to employ a material that is transparent to visible and/or UV light. For flexible substrates, materials of interest include: nylon, both modified and unmodified, nitrocellulose, polypropylene, and the like, where a nylon membrane, as well as derivatives thereof, may be particularly useful in this embodiment. For rigid substrates, specific materials of interest include: glass; plastics (for example, polytetrafluoroethylene, polypropylene, polystyrene, polycarbonate, and blends thereof, and the like); metals (for example, gold, platinum, and the like).
- The substrate surface onto which the polynucleotide compositions or other moieties is deposited may be smooth or substantially planar, or have irregularities, such as depressions or elevations. The surface may be modified with one or more different layers of compounds that serve to modify the properties of the surface in a desirable manner. Such modification layers, when present, will generally range in thickness from a monomolecular thickness to about 1 mm, usually from a monomolecular thickness to about 0.1 mm and more usually from a monomolecular thickness to about 0.001 mm. Modification layers of interest include: inorganic and organic layers such as metals, metal oxides, polymers, small organic molecules and the like. Polymeric layers of interest include layers of: peptides, proteins, polynucleic acids or mimetics thereof (for example, peptide nucleic acids and the like); polysaccharides, phospholipids, polyurethanes, polyesters, polycarbonates, polyureas, polyamides, polyethyleneamines, polyarylene sulfides, polysiloxanes, polyimides, polyacetates, and the like, where the polymers may be hetero- or homopolymeric, and may or may not have separate functional moieties attached thereto (for example, conjugated), It will be appreciated from the above description that the apparatus and method, by including a
venturi 80, provides a simple way of providing negative or positive backpressure in a pulse type fluid dispensing head while conveniently isolating sensitive reagents to contaminants (such as moisture). It will also be appreciated that variations and modifications of the above described embodiments of the invention are, of course, possible. Accordingly, the present invention is not limited to such embodiments.
Claims (35)
1. A method of fabricating an array of biopolymers on a substrate using a biopolymer or biomonomer fluid, and using a fluid dispensing head having:
at least one jet which can dispense droplets onto a substrate, the jet including a chamber with an orifice, and including an ejector which, when activated, causes a droplet to be ejected from the orifice;
the method comprising:
(a) positioning the head with the orifice facing the substrate;
(b) dispensing multiple droplets of the biopolymer or biomonomer fluid from the head so as to form an array of droplets on the substrate;
(c) directing a gas flow through a venturi which has a throat opening communicating with the dispensing head chamber;
(d) varying gas flow resistance on an outlet side of the venturi, to alter the chamber pressure.
2. A method according to wherein gas flow resistance on the venturi outlet side is adjusted to alter the chamber pressure before or after step (b).
claim 1
3. A method according to wherein a negative spotting pressure is provided from the venturi throat opening to the head chamber during dispensing of the droplets, and wherein the gas flow resistance of the venturi outlet side is adjusted to provide a positive chamber pressure.
claim 1
4. A method according to additionally comprising loading the chamber with fluid from a direction behind the orifice and wherein, following loading, the gas flow resistance of the venturi outlet side is increased to provide a positive priming pressure to the chamber so as to assist in priming the at least one jet.
claim 1
5. A method according to wherein the gas flow resistance of the venturi outlet side is increased to provide a positive purging pressure to the chamber so as to purge any fluid remaining in the chamber through the orifice.
claim 1
6. A method according to additionally comprising adding a purge fluid to the chamber prior to providing the purging pressure to the chamber.
claim 3
7. A method according to wherein the gas directed through the venturi is an anhydrous compressed gas.
claim 1
8. A method according to wherein each fluid droplet has a volume of from 0.1 to 1000 pL.
claim 1
9. A method according to additionally comprising altering the gas flow rate through the venturi by adjusting a valve on an inlet side of the venturi.
claim 1
10. A method according to additionally comprising:
claim 1
providing a spotting pressure in the chamber from the venturi throat opening while the head is facing the substrate so as to retain fluid in the chamber in the absence of the ejector being activated;
and wherein the venturi outlet side gas flow resistance is increased to provide a purging pressure which is greater than the spotting pressure and sufficiently positive so as to purge fluid remaining in the chamber through the orifice.
11. A method according to additionally comprising adding a purge fluid to the head prior to providing the purging pressure to the head.
claim 10
12. A method according to additionally comprising, prior to step (a):
claim 1
positioning the head facing a load station spaced from the substrate, with the orifice adjacent and facing the biomonomer or biopolymer fluid, and
providing a loading pressure in the chamber from the venturi throat opening while the head is facing the load station, which is sufficiently negative such that the fluid is drawn into the chamber through the orifice;
and wherein the gas flow rate through the venturi is adjusted to provide a spotting pressure while dispensing droplets from the head, which spotting pressure is higher than the loading pressure.
13. A method according to wherein the gas flow rate is adjusted to provide the spotting pressure by adjusting a valve on an inlet side of the venturi.
claim 12
14. A method according to additionally comprising:
claim 1
positioning the head facing a load station with the orifice adjacent and facing the biomonomer or biopolymer fluid;
providing a loading pressure in the chamber from the venturi throat opening while the head is facing the load station, which is sufficiently negative such that the fluid is drawn into the chamber through the orifice;
positioning the head facing a purge station which is spaced from the substrate;
and wherein:
the gas flow rate through the venturi is adjusted by adjusting a valve on an inlet side of the venturi to provide a spotting pressure while dispensing droplets from the head, which spotting pressure is higher than the loading pressure; and
following dispensing of droplets, the venturi outlet side gas flow resistance is increased to provide a purging pressure in the chamber while the head is facing the purge station which is sufficiently positive such that fluid in the chamber is purged through the orifice.
15. A method of fabricating an array of biopolymers on a substrate using a biopolymer or biomonomer fluid, and using a fluid dispensing head having:
at least one jet which can dispense droplets onto a substrate, the jet including a chamber with an orifice, and including an ejector which, when activated, causes a droplet to be ejected from the orifice;
the method comprising:
(a) positioning the head with the orifice facing the substrate;
(b) dispensing multiple droplets of the biopolymer or biomonomer fluid from the head so as to form an array of droplets on the substrate;
(c) directing a flow of anhydrous gas through a venturi which has a throat opening communicating with the dispensing head chamber.
16. A method according to additionally comprising providing a spotting pressure in the chamber from the venturi throat opening while the head is facing the substrate so as to retain fluid in the chamber in the absence of the ejector being activated.
claim 15
17. A method according to additionally comprising:
claim 15
positioning the head facing a purge station which is spaced from the substrate; and
providing a purging pressure in the chamber from the venturi throat opening so as to purge fluid remaining in the chamber through the orifice.
18. A method according to additionally comprising, prior to step (a):
claim 15
positioning the head facing a load station which is spaced from the substrate, with the orifice adjacent and facing the biomonomer or biopolymer fluid, and
providing a loading pressure in the chamber from the venturi throat opening while the head is facing the load station, which is sufficiently negative such that the fluid is drawn into the chamber through the orifice.
19. A method according to wherein the head has multiple pulse jets with orifices on a common front face of the head.
claim 15
20. A method according to wherein fluid is simultaneously drawn in through the orifice of the multiple jets when the head is facing the loading station.
claim 18
21. An apparatus for fabricating an array of biopolymers on a substrate, comprising:
(a) a substrate station on which the substrate can be mounted;
(b) a dispensing head having:
at least one jet which can dispense droplets onto a substrate, the jet including a chamber with an orifice, and including an ejector which, when activated, causes a droplet to be ejected from the orifice;
(c) a positioning system to move at least one of the dispensing head and mounted substrate with respect to the other so that multiple droplets dispensed from the head onto the substrate will form an array thereon;
(d) a venturi having an inlet and an outlet, and a throat opening communicating with the chamber; and
(e) an adjustable venturi outlet control valve communicating with the venturi outlet so that selectable pressure can be applied to the head chamber by adjustment of the outlet control valve.
22. An apparatus according to wherein the venturi outlet control valve can be adjusted to apply positive or negative pressure to the dispensing head chamber.
claim 21
23. An apparatus according to additionally comprising a load station spaced from the substrate station to retain multiple biopolymer or polymerizable biomonomer reagent solutions for loading into the head.
claim 21
24. An apparatus according to wherein each fluid droplet has a volume of from 0.1 to 1000 pL.
claim 21
25. An apparatus according to additionally comprising a source of anhydrous compressed gas communicating with the venturi inlet.
claim 21
26. An apparatus according to wherein the venturi outlet control valve is adjustable to provide selectable different negative pressures to the chamber.
claim 21
27. An apparatus according to additionally comprising:
claim 21
a purge station spaced from the substrate station; and
a control processor which operates the positioning system to selectively position the head facing any one of the stations, and which adjusts the venturi outlet control valve to a restricted position so as to provide, when the head is facing the purge station, a positive pressure to the chamber from the venturi throat opening to purge fluid from the dispensing head.
28. An apparatus according to additionally comprising:
claim 21
a load station spaced from the substrate station; a control processor which operates the positioning system to selectively position the head facing any one of the stations, and which adjusts the venturi outlet control valve so as to provide, when the head is facing the load station, a negative pressure to the chamber from the venturi throat opening to draw fluid positioned adjacent the orifice into the chamber through the orifice.
29. An apparatus according to additionally comprising:
claim 21
a load station and a purge station which are spaced from the substrate station; and a control processor which operates the positioning system to selectively position the dispensing head facing any one of the stations;
and wherein:
the control processor adjusts the venturi outlet control valve to apply negative pressure to the chamber from the venturi throat opening when the dispensing head is facing the load station, so as to draw fluid positioned adjacent the orifice into the chamber through the orifice; and
the control processor adjusts the venturi outlet control valve so that positive pressure is applied to the chamber from the venturi throat opening when the dispensing head is facing the purge location, to force out any fluid in the head.
30. An apparatus according to wherein the head has multiple pulse jets with orifices on a common front face of the head.
claim 21
31. An apparatus for fabricating an array of biopolymers on a substrate, comprising:
(a) a substrate station at which the substrate can be mounted;
(b) a dispensing head having:
at least one jet which can dispense droplets onto a substrate, the jet including a chamber with an orifice, and including an ejector which, when activated, causes a droplet to be ejected from the orifice;
(c) a positioning system to move at least one of the dispensing head and mounted substrate with respect to the other so that multiple droplets dispensed from the head onto the substrate will form an array thereon;
(d) a venturi having an inlet and an outlet, and a throat opening communicating with the chamber; and
(e) a source of anhydrous compressed gas communicating with the venturi pressurized gas inlet.
32. An apparatus according to additionally comprising a valve to adjust the gas flow rate through the venturi.
claim 31
33. An apparatus according to wherein the valve is on an inlet side of the venturi.
claim 31
34. An apparatus according to wherein the head has multiple pulse jets with orifices on a common front face of the head.
claim 31
35. An apparatus for fabricating an array of biopolymers on a substrate, comprising:
(a) a substrate station at which the substrate can be mounted;
(b) a dispensing head having:
at least one jet which can dispense droplets onto a substrate, the jet including a chamber with an orifice, and including an ejector which, when activated, causes a droplet to be ejected from the orifice;
(c) an enclosure about the substrate station and dispensing head so that the head and substrate station can be maintained in a controlled atmosphere;
(d) a positioning system to move at least one of the dispensing head and mounted substrate with respect to the other so that multiple droplets dispensed from the head onto the substrate will form an array thereon;
(d) a venturi having an inlet and an outlet, and a throat opening communicating with the chamber; and
(e) a valve to control gas flow rate through the venturi.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US09/820,476 US6372483B2 (en) | 1999-04-30 | 2001-03-28 | Preparation of biopolymer arrays |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US09/302,899 US6242266B1 (en) | 1999-04-30 | 1999-04-30 | Preparation of biopolymer arrays |
US09/820,476 US6372483B2 (en) | 1999-04-30 | 2001-03-28 | Preparation of biopolymer arrays |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/302,899 Division US6242266B1 (en) | 1999-04-30 | 1999-04-30 | Preparation of biopolymer arrays |
Publications (2)
Publication Number | Publication Date |
---|---|
US20010010916A1 true US20010010916A1 (en) | 2001-08-02 |
US6372483B2 US6372483B2 (en) | 2002-04-16 |
Family
ID=23169686
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/302,899 Expired - Fee Related US6242266B1 (en) | 1999-04-30 | 1999-04-30 | Preparation of biopolymer arrays |
US09/820,476 Expired - Fee Related US6372483B2 (en) | 1999-04-30 | 2001-03-28 | Preparation of biopolymer arrays |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/302,899 Expired - Fee Related US6242266B1 (en) | 1999-04-30 | 1999-04-30 | Preparation of biopolymer arrays |
Country Status (1)
Country | Link |
---|---|
US (2) | US6242266B1 (en) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20030012698A1 (en) * | 2001-05-01 | 2003-01-16 | Ngk Insulators, Ltd. | Method for manufacturing biochips |
US20030118717A1 (en) * | 2001-12-24 | 2003-06-26 | Peck Bill J. | Fluid exit in reaction chambers |
EP1462175A2 (en) * | 2003-03-24 | 2004-09-29 | Agilent Technologies, Inc. | Apparatus and methods for dispensing isolated droplet |
US20050214779A1 (en) * | 2004-03-29 | 2005-09-29 | Peck Bill J | Methods for in situ generation of nucleic acid arrays |
WO2005107949A1 (en) * | 2004-05-04 | 2005-11-17 | P.A.L.M. Microlaser Technologies Ag | Method and apparatus for creating an analysis arrangement comprising discrete, separate test zones used for performing biological, biochemical, or chemical analyses |
US10286399B2 (en) * | 2013-03-05 | 2019-05-14 | Touchlight IP Limited | Synthesis apparatus and method |
US11623237B2 (en) * | 2017-03-07 | 2023-04-11 | Tokyo Electron Limited | Droplet ejecting apparatus having correctable movement mechanism for workpiece table and droplet ejecting method |
Families Citing this family (197)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6955915B2 (en) * | 1989-06-07 | 2005-10-18 | Affymetrix, Inc. | Apparatus comprising polymers |
US6090555A (en) * | 1997-12-11 | 2000-07-18 | Affymetrix, Inc. | Scanned image alignment systems and methods |
US5959098A (en) | 1996-04-17 | 1999-09-28 | Affymetrix, Inc. | Substrate preparation process |
US6706875B1 (en) * | 1996-04-17 | 2004-03-16 | Affyemtrix, Inc. | Substrate preparation process |
US6323043B1 (en) * | 1999-04-30 | 2001-11-27 | Agilent Technologies, Inc. | Fabricating biopolymer arrays |
US7504213B2 (en) | 1999-07-09 | 2009-03-17 | Agilent Technologies, Inc. | Methods and apparatus for preparing arrays comprising features having degenerate biopolymers |
US6422684B1 (en) * | 1999-12-10 | 2002-07-23 | Sensant Corporation | Resonant cavity droplet ejector with localized ultrasonic excitation and method of making same |
US6420180B1 (en) * | 2000-01-26 | 2002-07-16 | Agilent Technologies, Inc. | Multiple pass deposition for chemical array fabrication |
US7731904B2 (en) * | 2000-09-19 | 2010-06-08 | Canon Kabushiki Kaisha | Method for making probe support and apparatus used for the method |
US20060141507A1 (en) * | 2000-09-27 | 2006-06-29 | Kronick Mel N | Manufacture and use of non-standard size microarray slides |
US6656740B1 (en) * | 2000-10-31 | 2003-12-02 | Agilent Technologies, Inc. | Pressure variation in array fabrication |
US6623700B1 (en) | 2000-11-22 | 2003-09-23 | Xerox Corporation | Level sense and control system for biofluid drop ejection devices |
US6713022B1 (en) * | 2000-11-22 | 2004-03-30 | Xerox Corporation | Devices for biofluid drop ejection |
US6740530B1 (en) | 2000-11-22 | 2004-05-25 | Xerox Corporation | Testing method and configurations for multi-ejector system |
US6861034B1 (en) | 2000-11-22 | 2005-03-01 | Xerox Corporation | Priming mechanisms for drop ejection devices |
US7166258B2 (en) * | 2001-01-31 | 2007-01-23 | Agilent Technologies, Inc. | Automation-optimized microarray package |
US7112305B2 (en) | 2001-01-31 | 2006-09-26 | Agilent Technologies, Inc. | Automation-optimized microarray package |
US20030170148A1 (en) * | 2001-01-31 | 2003-09-11 | Mcentee John F. | Reaction chamber roll pump |
TW490783B (en) * | 2001-05-22 | 2002-06-11 | Hi Max Optoelectronics Corp | Testing device and method built in the wafer scribe line |
JP4166690B2 (en) * | 2001-06-20 | 2008-10-15 | サイトノーム インコーポレーテッド | Droplet supply system |
US20030027219A1 (en) * | 2001-07-31 | 2003-02-06 | Ilsley Diane D. | Methods for depositing small volumes of protein fluids onto the surface of a substrate |
JP2003043041A (en) * | 2001-08-01 | 2003-02-13 | Canon Inc | Liquid discharge device and apparatus for manufacturing sample carrier |
US7666661B2 (en) | 2001-08-27 | 2010-02-23 | Platypus Technologies, Llc | Substrates, devices, and methods for quantitative liquid crystal assays |
JP4252451B2 (en) * | 2001-08-30 | 2009-04-08 | 浜松ホトニクス株式会社 | Liquid droplet forming method and liquid droplet forming apparatus |
US6852850B2 (en) * | 2001-10-31 | 2005-02-08 | Agilent Technologies, Inc. | Use of ionic liquids for fabrication of polynucleotide arrays |
US6858720B2 (en) * | 2001-10-31 | 2005-02-22 | Agilent Technologies, Inc. | Method of synthesizing polynucleotides using ionic liquids |
US6838888B2 (en) * | 2001-12-13 | 2005-01-04 | Agilent Technologies, Inc. | Flow cell humidity sensor system |
US6994429B1 (en) * | 2001-12-13 | 2006-02-07 | Agilent Technologies, Inc. | Printhead fluid supply system |
US6935727B2 (en) | 2001-12-18 | 2005-08-30 | Agilent Technologies, Inc. | Pulse jet print head assembly having multiple reservoirs and methods for use in the manufacture of biopolymeric arrays |
US7005293B2 (en) | 2001-12-18 | 2006-02-28 | Agilent Technologies, Inc. | Multiple axis printhead adjuster for non-contact fluid deposition devices |
US7141368B2 (en) * | 2002-01-30 | 2006-11-28 | Agilent Technologies, Inc. | Multi-directional deposition in array fabrication |
US20030143444A1 (en) * | 2002-01-31 | 2003-07-31 | Qin Liu | Fuel cell with fuel droplet fuel supply |
US20030156136A1 (en) * | 2002-02-15 | 2003-08-21 | Cattell Herbert F. | Method and system for visualization of results of feature extraction from molecular array data |
US6929951B2 (en) * | 2002-02-28 | 2005-08-16 | Agilent Technologies, Inc. | Method and system for molecular array scanner calibration |
US6770892B2 (en) | 2002-02-28 | 2004-08-03 | Agilent Technologies, Inc. | Method and system for automated focus-distance determination for molecular array scanners |
US6914229B2 (en) * | 2002-02-28 | 2005-07-05 | Agilent Technologies, Inc. | Signal offset for prevention of data clipping in a molecular array scanner |
US20060149503A1 (en) * | 2004-12-30 | 2006-07-06 | Minor James M | Methods and systems for fast least squares optimization for analysis of variance with covariants |
US7248973B2 (en) * | 2002-04-23 | 2007-07-24 | Agilent Technologies, Inc. | Metrics for characterizing chemical arrays based on analysis of variance (ANOVA) factors |
US20050143933A1 (en) * | 2002-04-23 | 2005-06-30 | James Minor | Analyzing and correcting biological assay data using a signal allocation model |
US20030216870A1 (en) * | 2002-05-07 | 2003-11-20 | Wolber Paul K. | Method and system for normalization of micro array data based on local normalization of rank-ordered, globally normalized data |
US7221785B2 (en) * | 2002-05-21 | 2007-05-22 | Agilent Technologies, Inc. | Method and system for measuring a molecular array background signal from a continuous background region of specified size |
US20030220746A1 (en) * | 2002-05-21 | 2003-11-27 | Srinka Ghosh | Method and system for computing and applying a global, multi-channel background correction to a feature-based data set obtained from scanning a molecular array |
US20050033525A1 (en) * | 2002-05-21 | 2005-02-10 | Corson John F. | Method and system for computing and applying a user-defined, global, multi-channel background correction to a feature-based data set obtained from reading a microarray |
CA2486812A1 (en) | 2002-05-22 | 2004-05-21 | Platypus Technologies, Llc | Substrates, devices, and methods for cellular assays |
US6806460B2 (en) * | 2002-05-31 | 2004-10-19 | Agilent Technologies, Inc. | Fluorescence detection with increased dynamic range |
US7078505B2 (en) * | 2002-06-06 | 2006-07-18 | Agilent Technologies, Inc. | Manufacture of arrays with varying deposition parameters |
US7371348B2 (en) * | 2002-06-14 | 2008-05-13 | Agilent Technologies | Multiple array format |
US20030232140A1 (en) * | 2002-06-14 | 2003-12-18 | Joseph Remick | Methods for reagent removal in flow chambers |
US20040009608A1 (en) * | 2002-07-10 | 2004-01-15 | Caren Michael P. | Arrays with positioning control |
US7153689B2 (en) * | 2002-08-01 | 2006-12-26 | Agilent Technologies, Inc. | Apparatus and methods for cleaning and priming droplet dispensing devices |
JP4168693B2 (en) * | 2002-08-02 | 2008-10-22 | セイコーエプソン株式会社 | Inkjet printing apparatus, liquid filling method for ink jet head, microarray manufacturing apparatus, and liquid filling method for discharge head thereof |
US20050216459A1 (en) * | 2002-08-08 | 2005-09-29 | Aditya Vailaya | Methods and systems, for ontological integration of disparate biological data |
US20050112689A1 (en) | 2003-04-04 | 2005-05-26 | Robert Kincaid | Systems and methods for statistically analyzing apparent CGH data anomalies and plotting same |
US20040038214A1 (en) * | 2002-08-23 | 2004-02-26 | Corson John F. | Method and system for reading a molecular array |
JP4112935B2 (en) * | 2002-09-30 | 2008-07-02 | 浜松ホトニクス株式会社 | Liquid droplet forming method and liquid droplet forming apparatus, and ink jet printing method and apparatus |
US20040081966A1 (en) * | 2002-10-25 | 2004-04-29 | Peck Bill J. | Chemical array linking layers |
US20040086869A1 (en) * | 2002-10-31 | 2004-05-06 | Schembri Carol T. | Device having multiple molecular arrays |
US7247337B1 (en) | 2002-12-16 | 2007-07-24 | Agilent Technologies, Inc. | Method and apparatus for microarray fabrication |
US20040152081A1 (en) * | 2003-01-31 | 2004-08-05 | Leproust Eric M. | Viscosity control during polynucleotide synthesis |
US20040151635A1 (en) * | 2003-01-31 | 2004-08-05 | Leproust Eric M. | Array fabrication using deposited drop splat size |
US7825929B2 (en) * | 2003-04-04 | 2010-11-02 | Agilent Technologies, Inc. | Systems, tools and methods for focus and context viewing of large collections of graphs |
US7750908B2 (en) * | 2003-04-04 | 2010-07-06 | Agilent Technologies, Inc. | Focus plus context viewing and manipulation of large collections of graphs |
US20040219663A1 (en) * | 2003-04-30 | 2004-11-04 | Page Robert D. | Biopolymer array fabrication using different drop deposition heads |
US20040241666A1 (en) * | 2003-05-30 | 2004-12-02 | Amorese Douglas A. | Ligand array assays that include an organic fluid wash step and compositions for practicing the same |
US20040243341A1 (en) * | 2003-05-30 | 2004-12-02 | Le Cocq Christian A. | Feature extraction methods and systems |
US20040241663A1 (en) * | 2003-05-30 | 2004-12-02 | Peck Bill J. | Ligand array processing methods that include a high surface tension fluid deposition step and compositions for practicing the same |
US20040263597A1 (en) * | 2003-06-24 | 2004-12-30 | Eastman Kodak Company | Apparatus and method of producing multiple spectral deposits from a mixture of a compressed fluid and a marking material |
US20050008932A1 (en) * | 2003-07-10 | 2005-01-13 | Plotkin Lawrence R. | Fluid supply device for electrochemical cell |
US20050064395A1 (en) | 2003-07-25 | 2005-03-24 | Platypus Technologies, Llc | Liquid crystal based analyte detection |
US7348144B2 (en) * | 2003-08-13 | 2008-03-25 | Agilent Technologies, Inc. | Methods and system for multi-drug treatment discovery |
US20050049796A1 (en) * | 2003-09-03 | 2005-03-03 | Webb Peter G. | Methods for encoding non-biological information on microarrays |
US20050048506A1 (en) * | 2003-09-03 | 2005-03-03 | Fredrick Joseph P. | Methods for encoding non-biological information on microarrays |
US7108979B2 (en) * | 2003-09-03 | 2006-09-19 | Agilent Technologies, Inc. | Methods to detect cross-contamination between samples contacted with a multi-array substrate |
JP2005095003A (en) * | 2003-09-03 | 2005-04-14 | Fuji Photo Film Co Ltd | Method for separating and purifying nucleic acid |
US7324677B2 (en) * | 2003-10-14 | 2008-01-29 | Agilent Technologies, Inc. | Feature quantitation methods and system |
US20050079102A1 (en) * | 2003-10-14 | 2005-04-14 | Staton Kenneth L. | Interrogation apparatus |
CA2545482A1 (en) | 2003-11-10 | 2005-05-26 | Platypus Technologies, Llc | Substrates, devices, and methods for cellular assays |
US7022157B2 (en) * | 2003-11-12 | 2006-04-04 | Agilent Technologies, Inc. | Devices and methods for performing array based assays |
US6985834B2 (en) * | 2003-12-16 | 2006-01-10 | Agilent Technologies, Inc. | Methods and system for comparing data values across multiple platforms |
AU2004308964B2 (en) | 2003-12-23 | 2010-09-16 | Arbor Vita Corporation | Antibodies for oncogenic strains of HPV and methods of their use |
US20050170378A1 (en) * | 2004-02-03 | 2005-08-04 | Yakhini Zohar H. | Methods and systems for joint analysis of array CGH data and gene expression data |
US20070031883A1 (en) * | 2004-03-04 | 2007-02-08 | Kincaid Robert H | Analyzing CGH data to identify aberrations |
US8321138B2 (en) * | 2005-07-29 | 2012-11-27 | Agilent Technologies, Inc. | Method of characterizing quality of hybridized CGH arrays |
US20050227221A1 (en) * | 2004-04-09 | 2005-10-13 | Minor James M | Methods and systems for evaluating and for comparing methods of testing tissue samples |
US7159959B2 (en) * | 2004-05-05 | 2007-01-09 | Agilent Technologies, Inc. | Methods and systems for detecting errors in printhead pattern data and for preventing erroneous printing |
US20060040287A1 (en) * | 2004-06-02 | 2006-02-23 | Corson John F | Method and system for quantifying random errors and any spatial-intensity trends present in microarray data sets |
US7302348B2 (en) | 2004-06-02 | 2007-11-27 | Agilent Technologies, Inc. | Method and system for quantifying and removing spatial-intensity trends in microarray data |
US20050281462A1 (en) * | 2004-06-16 | 2005-12-22 | Jayati Ghosh | System and method of automated processing of multiple microarray images |
US20050282174A1 (en) * | 2004-06-19 | 2005-12-22 | Webb Peter G | Methods and systems for selecting nucleic acid probes for microarrays |
US20060004527A1 (en) * | 2004-07-01 | 2006-01-05 | Sampas Nicholas M | Methods, systems and computer readable media for identifying dye-normalization probes |
EP1782321A4 (en) * | 2004-07-23 | 2009-11-04 | Univ North Carolina | Methods and materials for determining pain sensitivity and predicting and treating related disorders |
JP4302591B2 (en) | 2004-08-20 | 2009-07-29 | 浜松ホトニクス株式会社 | Droplet formation condition determination method, droplet volume measurement method, particle number measurement method, and droplet formation apparatus |
US20060046252A1 (en) * | 2004-08-30 | 2006-03-02 | Srinka Ghosh | Method and system for developing probes for dye normalization of microarray signal-intensity data |
US20060056671A1 (en) * | 2004-09-15 | 2006-03-16 | Jayati Ghosh | Automated feature extraction processes and systems |
US20060064246A1 (en) * | 2004-09-20 | 2006-03-23 | Medberry Scott L | Automated Processing of chemical arrays and systems therefore |
US7877213B2 (en) | 2004-09-23 | 2011-01-25 | Agilent Technologies, Inc. | System and methods for automated processing of multiple chemical arrays |
US20060063274A1 (en) * | 2004-09-23 | 2006-03-23 | Schremp Donald J | Methods for manufacturing and using chemical array calibration devices |
US9040305B2 (en) * | 2004-09-28 | 2015-05-26 | Singulex, Inc. | Method of analysis for determining a specific protein in blood samples using fluorescence spectrometry |
US8685711B2 (en) | 2004-09-28 | 2014-04-01 | Singulex, Inc. | Methods and compositions for highly sensitive detection of molecules |
US20060080043A1 (en) * | 2004-10-12 | 2006-04-13 | Sampas Nicholas M | Comparative genomic hybridization significance analysis using data smoothing with shaped response functions |
US20070099227A1 (en) * | 2004-10-12 | 2007-05-03 | Curry Bo U | Significance analysis using data smoothing with shaped response functions |
US20060078896A1 (en) * | 2004-10-12 | 2006-04-13 | Peck Bill J | Method, system, and computer program product to assess properties of a chemical array |
US20090123958A1 (en) * | 2004-11-08 | 2009-05-14 | Inkjet Technology Ltd. | Laboratory Devices, Methods and Systems Employing Acoustic Ejection Devices |
US20060110744A1 (en) | 2004-11-23 | 2006-05-25 | Sampas Nicolas M | Probe design methods and microarrays for comparative genomic hybridization and location analysis |
US20060173635A1 (en) * | 2005-02-01 | 2006-08-03 | Yakhini Zohar H | Analyzing and visualizing enrichment in DNA sequence alterations |
US20060247867A1 (en) * | 2005-04-29 | 2006-11-02 | Delenstarr Glenda C | Customized and dynamic association of probe type with feature extraction algorithms |
US20060249652A1 (en) * | 2005-05-03 | 2006-11-09 | Kyle Schleifer | Methods and systems for pixilation processing of precision, high-speed scanning |
US7396676B2 (en) | 2005-05-31 | 2008-07-08 | Agilent Technologies, Inc. | Evanescent wave sensor with attached ligand |
US20060282221A1 (en) * | 2005-06-09 | 2006-12-14 | Shah Manish M | Automatic array quality analysis |
US7333907B2 (en) * | 2005-07-29 | 2008-02-19 | Agilent Technologies, Inc. | System and methods for characterization of chemical arrays for quality control |
US7662572B2 (en) | 2005-08-25 | 2010-02-16 | Platypus Technologies, Llc. | Compositions and liquid crystals |
US20070067110A1 (en) * | 2005-09-21 | 2007-03-22 | Nelson Charles F Iii | Generation of negative controls for arrays |
US20070070132A1 (en) * | 2005-09-27 | 2007-03-29 | Fan-Cheung Sze | Inkjet delivery module |
US20070092882A1 (en) | 2005-10-21 | 2007-04-26 | Hui Wang | Analysis of microRNA |
US20070111218A1 (en) * | 2005-11-17 | 2007-05-17 | Ilsley Diane D | Label integrity verification of chemical array data |
US20070111219A1 (en) * | 2005-11-17 | 2007-05-17 | Minor James M | Label integrity verification of chemical array data |
EP1951910A4 (en) * | 2005-11-30 | 2010-02-03 | Univ North Carolina | Identification of genetic polymorphic variants associated with somatosensory disorders and methods of using the same |
US7807354B2 (en) | 2005-12-28 | 2010-10-05 | Agilent Technologies, Inc. | Low volume hybridization |
US9445025B2 (en) | 2006-01-27 | 2016-09-13 | Affymetrix, Inc. | System, method, and product for imaging probe arrays with small feature sizes |
US8055098B2 (en) * | 2006-01-27 | 2011-11-08 | Affymetrix, Inc. | System, method, and product for imaging probe arrays with small feature sizes |
US7838250B1 (en) | 2006-04-04 | 2010-11-23 | Singulex, Inc. | Highly sensitive system and methods for analysis of troponin |
EP2002260B1 (en) | 2006-04-04 | 2015-11-04 | Singulex, Inc. | Highly sensitive system and methods for analysis of troponin |
US20070238104A1 (en) * | 2006-04-07 | 2007-10-11 | Agilent Technologies, Inc. | Competitive oligonucleotides |
US8058055B2 (en) * | 2006-04-07 | 2011-11-15 | Agilent Technologies, Inc. | High resolution chromosomal mapping |
US20070255512A1 (en) * | 2006-04-28 | 2007-11-01 | Delenstarr Glenda C | Methods and systems for facilitating analysis of feature extraction outputs |
US20080058218A1 (en) * | 2006-05-03 | 2008-03-06 | Gordon David B | Arrays of compound probes and methods using the same |
US20070259345A1 (en) * | 2006-05-03 | 2007-11-08 | Agilent Technologies, Inc. | Target determination using compound probes |
US20070259346A1 (en) * | 2006-05-03 | 2007-11-08 | Agilent Technologies, Inc. | Analysis of arrays |
US20070259347A1 (en) * | 2006-05-03 | 2007-11-08 | Agilent Technologies, Inc. | Methods of increasing the effective probe densities of arrays |
US7674924B2 (en) | 2006-05-22 | 2010-03-09 | Third Wave Technologies, Inc. | Compositions, probes, and conjugates and uses thereof |
US20080171665A1 (en) * | 2006-05-24 | 2008-07-17 | Minor James M | Programmed changes in hybridization conditions to improve probe signal quality |
US20070275389A1 (en) * | 2006-05-24 | 2007-11-29 | Anniek De Witte | Array design facilitated by consideration of hybridization kinetics |
US7437249B2 (en) * | 2006-06-30 | 2008-10-14 | Agilent Technologies, Inc. | Methods and systems for detrending signal intensity data from chemical arrays |
CA2657576C (en) | 2006-07-14 | 2023-10-31 | The Regents Of The University Of California | Cancer biomarkers and methods of use thereof |
US7842499B2 (en) * | 2006-08-07 | 2010-11-30 | Platypus Technologies, Llc | Substrates, devices, and methods for cellular assays |
WO2008021071A2 (en) | 2006-08-07 | 2008-02-21 | Platypus Technologies, Llc | Substrates, devices, and methods for cellular assays |
KR101462431B1 (en) * | 2006-08-15 | 2014-11-17 | 에코신테틱스 리미티드 | Process for producing biopolymer nanoparticles |
EP1918838A3 (en) | 2006-10-05 | 2009-07-08 | Agilent Technologies, Inc. | Estimation of dynamic range of microarray DNA spike-in data by use of parametric curve-fitting |
US8077951B2 (en) * | 2006-10-12 | 2011-12-13 | Agilent Technologies, Inc. | Method and system for dynamic, automated detection of outlying feature and feature background regions during processing of data scanned from a chemical array |
US7881876B2 (en) * | 2006-10-12 | 2011-02-01 | Agilent Technologies, Inc. | Methods and systems for removing offset bias in chemical array data |
WO2008046056A1 (en) * | 2006-10-13 | 2008-04-17 | Welldoc Communications, Inc. | Reduction of nonspecific binding in nucleic acid assays and nucleic acid synthesis reactions |
US20080090236A1 (en) * | 2006-10-13 | 2008-04-17 | Yakhini Zohar H | Methods and systems for identifying tumor progression in comparative genomic hybridization data |
US20090149342A1 (en) * | 2006-10-13 | 2009-06-11 | Welldoc Communications | Method for reduction of nonspecific binding in nucleic acid assays, nucleic acid synthesis and multiplex amplification reactions |
US20080102453A1 (en) * | 2006-10-31 | 2008-05-01 | Jayati Ghosh | Methods and systems and analysis of CGH data |
US20080204501A1 (en) * | 2006-12-01 | 2008-08-28 | Shinichi Kurita | Inkjet print head pressure regulator |
US20080207960A1 (en) * | 2007-02-28 | 2008-08-28 | Eric Lin | Methods, compositions, and kits for post-hybridization processing of arrays |
US20080206851A1 (en) * | 2007-02-28 | 2008-08-28 | Dellinger Douglas J | Methods and compositions for RNA synthesis |
US20080231908A1 (en) * | 2007-03-19 | 2008-09-25 | Xerox Corporation | Methods and systems for classifying and prioritizing incoming facsimiles |
JP2010526555A (en) * | 2007-05-11 | 2010-08-05 | タフツ・メディカル・センター | Polynucleotides associated with age-related macular degeneration and methods for assessing patient risk |
ES2420973T3 (en) | 2007-07-25 | 2013-08-28 | University Of Louisville Research Foundation, Inc. | Micro-RNA associated with exosome as a diagnostic marker |
EP2193365A4 (en) * | 2007-08-20 | 2015-05-13 | Platypus Technologies Llc | Improved devices for cell assays |
WO2009029890A1 (en) * | 2007-08-29 | 2009-03-05 | Applied Materials, Inc. | Methods and apparatus for modular print head and adapter and rotation thereof with inkjet printer systems |
AU2008352940B2 (en) | 2007-12-19 | 2014-06-05 | Singulex, Inc. | Scanning analyzer for single molecule detection and methods of use |
JP2009191020A (en) * | 2008-02-14 | 2009-08-27 | Oki Electric Ind Co Ltd | Apparatus for synthesizing organic compound, and method for synthesizing organic compound |
EP2263085A4 (en) * | 2008-03-05 | 2011-07-06 | Singulex Inc | Methods and compositions for highly sensitive detection of molecules |
EP2283163B1 (en) | 2008-04-18 | 2015-07-08 | Tufts Medical Center | Polymorphisms associated with age-related macular degeneration and methods for evaluating patient risk |
US8178355B2 (en) | 2008-09-15 | 2012-05-15 | Platypus Technologies, Llc. | Detection of vapor phase compounds by changes in physical properties of a liquid crystal |
GB2464183A (en) * | 2008-09-19 | 2010-04-14 | Singulex Inc | Sandwich assay |
WO2010065750A1 (en) * | 2008-12-03 | 2010-06-10 | Ecosynthetix Inc. | Process for producing biopolymer nanoparticle biolatex compositions having enhanced performance and compositions based thereon |
US20100161607A1 (en) * | 2008-12-22 | 2010-06-24 | Jasjit Singh | System and method for analyzing genome data |
AU2010229767C1 (en) | 2009-03-27 | 2015-02-19 | The General Hospital Corporation | Markers related to age-related macular degeneration and uses therefor |
CA2762612A1 (en) | 2009-06-08 | 2010-12-16 | Singulex, Inc. | Highly sensitive biomarker panels |
US20110039735A1 (en) | 2009-08-13 | 2011-02-17 | Agilent Technologies, Inc. | Probe design for oligonucleotide fluorescence in situ hybridization (fish) |
KR20110092046A (en) * | 2010-02-08 | 2011-08-17 | 삼성전자주식회사 | Microarray probe synthesis device comprising capillary nozzle and method for synthesizing microarray probe using the same |
EP3508854A1 (en) | 2010-04-27 | 2019-07-10 | The Regents of The University of California | Cancer biomarkers and methods of use thereof |
WO2011140484A1 (en) | 2010-05-06 | 2011-11-10 | Singulex, Inc | Methods for diagnosing, staging, predicting risk for developing and identifying treatment responders for rheumatoid arthritis |
WO2012083072A2 (en) | 2010-12-16 | 2012-06-21 | Agilent Technologies, Inc. | LIGATION METHOD EMPLOYING EUKARYOTIC tRNA LIGASE |
US8478545B2 (en) | 2011-06-03 | 2013-07-02 | Agilent Technologies, Inc. | Identification of aberrant microarray features |
EP2682753A1 (en) * | 2012-05-08 | 2014-01-08 | Roche Diagniostics GmbH | Cartridge for Dispensing a Fluid Comprising a Reagent |
KR102062418B1 (en) | 2012-08-31 | 2020-01-03 | 도레이 카부시키가이샤 | Method for detecting target nucleic acid |
US20140274749A1 (en) | 2013-03-15 | 2014-09-18 | Affymetrix, Inc. | Systems and Methods for SNP Characterization and Identifying off Target Variants |
DK3030682T3 (en) | 2013-08-05 | 2020-09-14 | Twist Bioscience Corp | DE NOVO SYNTHESIZED GENE LIBRARIES |
US11543411B2 (en) | 2014-12-05 | 2023-01-03 | Prelude Corporation | DCIS recurrence and invasive breast cancer |
WO2016126987A1 (en) | 2015-02-04 | 2016-08-11 | Twist Bioscience Corporation | Compositions and methods for synthetic gene assembly |
US10669304B2 (en) | 2015-02-04 | 2020-06-02 | Twist Bioscience Corporation | Methods and devices for de novo oligonucleic acid assembly |
WO2016172377A1 (en) | 2015-04-21 | 2016-10-27 | Twist Bioscience Corporation | Devices and methods for oligonucleic acid library synthesis |
WO2017049231A1 (en) | 2015-09-18 | 2017-03-23 | Twist Bioscience Corporation | Oligonucleic acid variant libraries and synthesis thereof |
US11512347B2 (en) | 2015-09-22 | 2022-11-29 | Twist Bioscience Corporation | Flexible substrates for nucleic acid synthesis |
CA3006867A1 (en) | 2015-12-01 | 2017-06-08 | Twist Bioscience Corporation | Functionalized surfaces and preparation thereof |
EP3500672A4 (en) | 2016-08-22 | 2020-05-20 | Twist Bioscience Corporation | De novo synthesized nucleic acid libraries |
US10417457B2 (en) | 2016-09-21 | 2019-09-17 | Twist Bioscience Corporation | Nucleic acid based data storage |
US10538796B2 (en) | 2016-10-13 | 2020-01-21 | Agilent Technologies, Inc. | On-array ligation assembly |
EP3554514A4 (en) | 2016-12-16 | 2020-08-05 | Twist Bioscience Corporation | Variant libraries of the immunological synapse and synthesis thereof |
EP3586255A4 (en) | 2017-02-22 | 2021-03-31 | Twist Bioscience Corporation | Nucleic acid based data storage |
WO2018170169A1 (en) | 2017-03-15 | 2018-09-20 | Twist Bioscience Corporation | Variant libraries of the immunological synapse and synthesis thereof |
AU2018284227A1 (en) | 2017-06-12 | 2020-01-30 | Twist Bioscience Corporation | Methods for seamless nucleic acid assembly |
WO2018231864A1 (en) | 2017-06-12 | 2018-12-20 | Twist Bioscience Corporation | Methods for seamless nucleic acid assembly |
KR20200047706A (en) | 2017-09-11 | 2020-05-07 | 트위스트 바이오사이언스 코포레이션 | GPCR binding protein and method for synthesis thereof |
SG11202003574TA (en) | 2017-10-20 | 2020-05-28 | Twist Bioscience Corp | Heated nanowells for polynucleotide synthesis |
CA3088911A1 (en) | 2018-01-04 | 2019-07-11 | Twist Bioscience Corporation | Dna-based digital information storage |
WO2019222706A1 (en) | 2018-05-18 | 2019-11-21 | Twist Bioscience Corporation | Polynucleotides, reagents, and methods for nucleic acid hybridization |
WO2020056338A1 (en) | 2018-09-14 | 2020-03-19 | Prelude Corporation | Method of selection for treatment of subjects at risk of invasive breast cancer |
US11492727B2 (en) | 2019-02-26 | 2022-11-08 | Twist Bioscience Corporation | Variant nucleic acid libraries for GLP1 receptor |
WO2020176680A1 (en) | 2019-02-26 | 2020-09-03 | Twist Bioscience Corporation | Variant nucleic acid libraries for antibody optimization |
CN114729342A (en) | 2019-06-21 | 2022-07-08 | 特韦斯特生物科学公司 | Barcode-based nucleic acid sequence assembly |
KR102231063B1 (en) * | 2019-11-06 | 2021-03-23 | 주식회사 엘지화학 | Device of manufacturing for chip of diagnosing allergy |
WO2023249835A1 (en) | 2022-06-23 | 2023-12-28 | Nanopec, Inc. | Apparatus and processes for high throughput automation of synthetic dna and rna on nanostructured ceramic films |
Family Cites Families (22)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3974508A (en) | 1974-12-16 | 1976-08-10 | Gould Inc. | Air purging system for a pulsed droplet ejecting system |
GB1527444A (en) | 1977-03-01 | 1978-10-04 | Itt Creed | Ink drop printhead |
US4314264A (en) | 1980-08-15 | 1982-02-02 | The Mead Corporation | Ink supply system for an ink jet printer |
US4399446A (en) | 1982-01-18 | 1983-08-16 | The Mead Corporation | Ink supply system for an ink jet printer |
IT1162919B (en) | 1983-07-20 | 1987-04-01 | Olivetti & Co Spa | INK JET WRITING DEVICE PARTICULARLY FOR HIGH SPEED PRINTERS |
US4614948A (en) * | 1985-04-12 | 1986-09-30 | Eastman Kodak Company | Ink circulation system for continuous ink jet printing apparatus |
US4607261A (en) | 1985-04-12 | 1986-08-19 | Eastman Kodak Company | Ink supply cartridge and cooperative ink circulation system of continuous ink jet printer |
US4771295B1 (en) | 1986-07-01 | 1995-08-01 | Hewlett Packard Co | Thermal ink jet pen body construction having improved ink storage and feed capability |
US4877745A (en) | 1986-11-17 | 1989-10-31 | Abbott Laboratories | Apparatus and process for reagent fluid dispensing and printing |
US4875055A (en) | 1988-11-21 | 1989-10-17 | Eastman Kodak Company | Simplified multicolor fluid system for continuous ink jet printer |
US4931811A (en) | 1989-01-31 | 1990-06-05 | Hewlett-Packard Company | Thermal ink jet pen having a feedtube with improved sizing and operational with a minimum of depriming |
US4929969A (en) | 1989-08-25 | 1990-05-29 | Eastman Kodak Company | Ink supply construction and printing method for drop-on-demand ink jet printing |
US5121132A (en) | 1989-09-29 | 1992-06-09 | Hewlett-Packard Company | Ink delivery system for printers |
KR970009104B1 (en) | 1992-09-30 | 1997-06-05 | Samsung Electronics Co Ltd | Recording method and apparatus of ink-jet printer using electric viscous fluid |
US6015880A (en) | 1994-03-16 | 2000-01-18 | California Institute Of Technology | Method and substrate for performing multiple sequential reactions on a matrix |
US5539952A (en) | 1994-08-22 | 1996-07-30 | Hayes; Thomas | Fluid handling apparatus for maintaining lithographic presses |
US5563639A (en) | 1994-09-30 | 1996-10-08 | Hewlett-Packard Company | Venturi spittoon system to control inkjet aerosol |
US5658802A (en) | 1995-09-07 | 1997-08-19 | Microfab Technologies, Inc. | Method and apparatus for making miniaturized diagnostic arrays |
US5681757A (en) | 1996-04-29 | 1997-10-28 | Microfab Technologies, Inc. | Process for dispensing semiconductor die-bond adhesive using a printhead having a microjet array and the product produced by the process |
US5981733A (en) | 1996-09-16 | 1999-11-09 | Incyte Pharmaceuticals, Inc. | Apparatus for the chemical synthesis of molecular arrays |
US5874554A (en) | 1996-12-13 | 1999-02-23 | Incyte Pharmaceuticals, Inc. | Methods and solvent vehicles for reagent delivery in oligonucleotide synthesis using automated pulse jetting devices |
WO1998041531A2 (en) | 1997-03-20 | 1998-09-24 | University Of Washington | Solvent for biopolymer synthesis, solvent microdroplets and methods of use |
-
1999
- 1999-04-30 US US09/302,899 patent/US6242266B1/en not_active Expired - Fee Related
-
2001
- 2001-03-28 US US09/820,476 patent/US6372483B2/en not_active Expired - Fee Related
Cited By (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1385000A4 (en) * | 2001-05-01 | 2006-04-19 | Ngk Insulators Ltd | Method for making biochip |
EP1385000A1 (en) * | 2001-05-01 | 2004-01-28 | Ngk Insulators, Ltd. | Method for making biochip |
US20030012698A1 (en) * | 2001-05-01 | 2003-01-16 | Ngk Insulators, Ltd. | Method for manufacturing biochips |
US7160512B2 (en) | 2001-05-01 | 2007-01-09 | Ngk Insulators, Ltd. | Method for manufacturing biochips |
US20030118717A1 (en) * | 2001-12-24 | 2003-06-26 | Peck Bill J. | Fluid exit in reaction chambers |
US6846454B2 (en) * | 2001-12-24 | 2005-01-25 | Agilent Technologies, Inc. | Fluid exit in reaction chambers |
US20050201896A1 (en) * | 2001-12-24 | 2005-09-15 | Peck Bill J. | Fluid exit in reaction chambers |
EP1462175A2 (en) * | 2003-03-24 | 2004-09-29 | Agilent Technologies, Inc. | Apparatus and methods for dispensing isolated droplet |
US20040191777A1 (en) * | 2003-03-24 | 2004-09-30 | Peck Bill J. | Apparatus and methods for isolating droplet dispensing devices |
EP1462175A3 (en) * | 2003-03-24 | 2005-08-03 | Agilent Technologies, Inc. | Apparatus and methods for dispensing isolated droplet |
US20050214779A1 (en) * | 2004-03-29 | 2005-09-29 | Peck Bill J | Methods for in situ generation of nucleic acid arrays |
EP1595962A3 (en) * | 2004-03-29 | 2005-11-23 | Agilent Technologies, Inc. | Methods for in situ generation of nucleic acid arrays |
US20060078927A1 (en) * | 2004-03-29 | 2006-04-13 | Peck Bill J | Methods for in situ generation of nucleic acid molecules |
EP1595962A2 (en) * | 2004-03-29 | 2005-11-16 | Agilent Technologies, Inc. | Methods for in situ generation of nucleic acid arrays |
WO2005107949A1 (en) * | 2004-05-04 | 2005-11-17 | P.A.L.M. Microlaser Technologies Ag | Method and apparatus for creating an analysis arrangement comprising discrete, separate test zones used for performing biological, biochemical, or chemical analyses |
US10286399B2 (en) * | 2013-03-05 | 2019-05-14 | Touchlight IP Limited | Synthesis apparatus and method |
US11623237B2 (en) * | 2017-03-07 | 2023-04-11 | Tokyo Electron Limited | Droplet ejecting apparatus having correctable movement mechanism for workpiece table and droplet ejecting method |
Also Published As
Publication number | Publication date |
---|---|
US6372483B2 (en) | 2002-04-16 |
US6242266B1 (en) | 2001-06-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US6372483B2 (en) | Preparation of biopolymer arrays | |
US7282332B2 (en) | Fabricating biopolymer arrays | |
US6461812B2 (en) | Method and multiple reservoir apparatus for fabrication of biomolecular arrays | |
US6900048B2 (en) | Biopolymer arrays and their fabrication | |
US6943036B2 (en) | Error detection in chemical array fabrication | |
US6890760B1 (en) | Array fabrication | |
US6599693B1 (en) | Array fabrication | |
US6656740B1 (en) | Pressure variation in array fabrication | |
US6420180B1 (en) | Multiple pass deposition for chemical array fabrication | |
US7101508B2 (en) | Chemical array fabrication errors | |
US8084245B2 (en) | Apparatus and method for polymer synthesis using arrays | |
US6946285B2 (en) | Arrays with elongated features | |
US20040018635A1 (en) | Fabricating arrays with drop velocity control | |
US20020106812A1 (en) | Fluid drop dispensing | |
US20090170729A1 (en) | Method and a machine for ex situ fabrication of low and medium integration biochip arrays | |
US20040009608A1 (en) | Arrays with positioning control | |
JP2002286732A (en) | Liquid discharge device used in manufacturing probe carrier, method of manufacturing probe carrier, and device of manufacturing probe carrier |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
FPAY | Fee payment |
Year of fee payment: 4 |
|
REMI | Maintenance fee reminder mailed | ||
LAPS | Lapse for failure to pay maintenance fees | ||
STCH | Information on status: patent discontinuation |
Free format text: PATENT EXPIRED DUE TO NONPAYMENT OF MAINTENANCE FEES UNDER 37 CFR 1.362 |
|
FP | Lapsed due to failure to pay maintenance fee |
Effective date: 20100416 |