EP2599548A1 - Sample holder for positioning an organic, biological and/or medical sample - Google Patents

Abstract

Sample holder comprises a structural element (103), which is designed such that an introduced organic, biological and/or medical sample in a sample holder is placed in a desired partial region, preferably in a desired surface area (104) of the sample holder. An independent claim is also included for positioning the organic, biological and/or medical sample in the desired surface region of the sample holder, comprising providing the sample holder as above, introducing the sample into the sample holder, and moving the sample holder such that the sample is placed in the desired surface area of the sample holder.

Description

The invention relates to a method for positioning an organic, biological and / or medical sample in a desired surface area of a sample carrier. In particular, the invention relates to a method for positioning a sample by means of a magnetic device.

Particularly in the fields of cell biology and medicine, sample carriers are used for the examination of organic, biological and / or medical samples. In most experiments, it is advantageous if the sample can be accurately positioned. This allows an efficient execution of the experiments, an increased comparability of several experiments and a simplification of the evaluation.

Frequently, when a sample carrier is filled, a random arrangement of the sample occurs. The positioning of the sample depends mostly on the geometry of the sample carrier and on the type of filling. In certain cases, the geometry of a sample carrier is designed to be counterproductive to a desired positioning of the sample.

It is therefore an object of the invention to provide a method for positioning an organic, biological and / or medical sample, which allows to position the sample in a desired surface area of the sample carrier.

This problem is solved by a method according to claim 1.

• Verbinden der Probe mit einem oder mehreren magnetischen, insbesondere paramagnetischen Partikeln,
• Anordnen der Magnetvorrichtung relativ zum Probenträger, so dass in einem vorherbestimmten Bereich des Probenträgers eine gewünschte Magnetfeldanordnung bereitgestellt wird,
• Einbringen der Probe in den Probenträger, und
• Anordnen der Probe in dem gewünschten Oberflächenbereich mit Hilfe der Magnetvorrichtung.

The method according to the invention for positioning an organic, biological and / or medical sample in a desired surface area of a sample carrier, wherein a magnetic device is provided, comprises the steps:

• Connecting the sample to one or more magnetic, in particular paramagnetic, particles
• Arranging the magnetic device relative to the sample carrier such that a desired magnetic field arrangement is provided in a predetermined region of the sample carrier,
• Introducing the sample into the sample carrier, and
• Arranging the sample in the desired surface area by means of the magnetic device.

This method enables a precise positioning of the sample in a desired surface area of the sample carrier.

The organic, biological and / or medical sample may be a biological cell. In particular, the method may be performed for a plurality of cells. As a result, a desired cell distribution in a desired surface area of the sample carrier can be achieved. In this case, the cells can be introduced in the form of a suspension in the sample carrier. The sample may also be a microorganism or DNA.

The sample carrier may comprise a plastic, in particular COC (cycloolefin copolymer), COP (cycloolefin polymer), PS (polystyrene), PC (polycarbonate) or PMMA (polymethyl methacrylate). The sample carrier may be formed as an injection molded part. The sample carrier may comprise a bottom plate, in particular wherein the sample carrier rests on the bottom plate during operation, and wherein the bottom plate may comprise a plastic and / or glass. The bottom plate can be thin, for example between 1 .mu.m and 300 .mu.m. This allows high-resolution microscopy through the bottom plate.

The sample carrier may be dimensioned such that the volume of a cavity is in the range of 5 .mu.l to 1000 .mu.l, in particular between 100 .mu.l and 500 .mu.l. Thus, the sample carrier for Mikrofluiduntersuchungen is usable.

The sample carrier may comprise a cover plate, wherein the cover plate with the bottom plate is liquid-tight, in particular directly connected.

The bottom plate and / or top plate may have a predetermined intrinsic fluorescence, which is in particular less than or equal to the intrinsic fluorescence of COC or COP or a conventional cover glass, and / or a predetermined refractive index, in particular> 1.2 and / or <1.7. In particular, the intrinsic fluorescence may be less than or equal to the intrinsic fluorescence of a conventional coverslip (for example, pure white glass of hydrolytic class 1 (such as Menzel coverslip, in particular having a starch # 1.5.) In particular, the predetermined refractive index may be> 1.2 and / or <1.7, with such a high-quality optical material microscopy investigations can advantageously be carried out, for example, the birefringence can be so low that DIC (Differential Interference Contrast) is possible.

In particular, the bottom plate and / or cover plate can be antireflective in a frequency range of the electromagnetic radiation used for microscopy. This allows the Transmission through the bottom plate and / or cover plate can be increased so that single molecule measurements are possible with the help of fluorescence.

The sample carrier may comprise at least one surface region for arranging a sample, in particular wherein the surface region is arranged on the bottom plate. The sample carrier may include a cavity for receiving a sample. At least one opening for filling and / or emptying the cavity with the sample and / or a liquid can lead into the cavity. The cavity may be formed by recesses in the cover plate and / or in the bottom plate.

By connecting the sample to one or more magnetic, in particular paramagnetic particles, samples which do not have their own magnetic moment can also be positioned with the aid of the magnetic device. If the sample is a living cell, the magnetic particles may be made of a material that does not have a toxic effect on the cell.

Arranging the sample may include aligning the sample in the desired magnetic field arrangement. In particular, the sample can align itself by the magnetic force effect of the desired magnetic field arrangement. In particular, the sample can move as a result of the magnetic force action and thereby an arrangement of the sample can be achieved.

The placement of the sample may include moving the magnetic device relative to the sample carrier. In this case, a sample which has been introduced in a predetermined area of the sample carrier may be arranged in a desired surface area. This may be particularly advantageous if the desired surface area is located in an inaccessible or hard to reach area of the sample carrier. In particular, the sample can align in the desired magnetic field arrangement and thereafter be moved by moving the magnetic device by magnetic force action in the desired surface area.

The predetermined area of the sample carrier may comprise the desired surface area. In this case, the placement of the sample can only be done by aligning the sample in the desired magnetic field arrangement.

The desired magnetic field arrangement may have a magnetic field strength, a magnetic force flux and / or a magnetic field line distribution. In particular, the magnetic device can provide a magnetic field, wherein the magnetic field, in particular the desired arrangement of the magnetic field or desired magnetic field arrangement in predetermined range can be characterized by a magnetic field strength, a magnetic flux and / or a magnetic field line distribution.

The magnetic field strength corresponds to a vectorial quantity and is also referred to as magnetic induction or magnetic flux density. The magnetic field strength is proportional to the magnetic excitation.

The predetermined region of the sample carrier may comprise a surface region of the sample carrier and the amount of the magnetic field strength in the predetermined region, in particular in the surface region, may have at least one local extremum, in particular a local maximum, and / or at least one saddle point. In this way, the sample can be moved away by magnetic force to the local extremum or away from the local extremum. By selecting the strength and / or the position of the local extremum, a targeted arrangement of the sample is possible.

For example, the surface area may comprise a partial area in which the magnetic field lines condense. In other words, in this subarea, the desired magnetic field arrangement has a local maximum in the amount of the magnetic field strength. This also means that the magnetic flux through the surface in this subregion can have a local maximum.

The magnetic field strength of the desired magnetic field arrangement may have a magnetic field component parallel and / or perpendicular to the surface area at any point or in a plurality of points of the predetermined range. Thereby, the sample can be moved parallel and / or perpendicular to the surface area by magnetic force. In particular, in combination with a local extremum can be achieved in this way, the placement of the sample by aligning the sample in the desired magnetic field arrangement.

The magnetic device may provide a dipole field or a quadrupole field. In particular, the magnetic device may also provide a combination of multiple dipole fields and / or quadrupole fields. In combination with the relative position of the sample carrier relative to the magnetic device, the magnetic field arrangement can thereby be varied or predetermined in the predetermined range.

The desired magnetic field arrangement, in particular its magnetic field line distribution, may be radially symmetrical with respect to a predetermined axis. This can be achieved, for example, when the magnetic device provides a dipole field. In particular, the be predetermined axis perpendicular to the surface area of the sample carrier. This allows the sample to be arranged in a radially symmetric surface area.

The bonding of the sample to one or more magnetic, in particular paramagnetic, particles may include adhering a particle to the surface of the sample and / or picking up or introducing a particle into the sample. In particular, if the sample does not have its own magnetic moment, by connecting the sample with a magnetic particle, the placement of the sample can be realized by means of a magnetic force effect of the desired magnetic field arrangement on the magnetic particle.

If the sample corresponds to a biological cell, the particle can be taken up by the cell. In this case, the magnetic particle is smaller than the cell, in particular, the volume and the maximum spatial extent of the magnetic particle is smaller than the volume and the maximum spatial extent of the cell. The magnetic, in particular paramagnetic, particle can be attached to the surface of the cell. This can be achieved by positively charged end groups. The particle can then be taken up (phagocytosed) by the cell. The particle can be stored in particular in vesicles in the cytosol.

The magnetic, in particular paramagnetic, particles can be coated with a polymer matrix, in particular wherein the polymer matrix is provided with a coating which can adhere to a surface of the sample. In this way, a particle can be connected to the surface of the sample. In this case, the particle may be larger than when it is to be introduced into the sample. In particular, if the sample corresponds to a biological cell, for example, a particle one-fiftieth of cell size may be used. The coating of the polymer matrix may comprise surface proteins, in particular CD molecules or activated tosyl groups. The coating can be chosen such that the particle can adhere to a desired cell type, especially only to the desired cell type.

The magnetic device may comprise a permanent magnet and / or an electromagnet. The permanent magnet may in particular be a neodymium iron boring magnet. As a result, a particularly high field strength can be achieved. For example, the maximum amount of magnetic field strength may be between 0.5 Tesla and 1.4 Tesla.

An electromagnet may include a coil having one or more turns. In particular, the electromagnet may comprise an iron core.

The magnetic device may comprise at least one tip, in particular wherein the tip comprises a magnetic, in particular ferromagnetic, material. As a result, in the region of the tip, a high density of magnetic field lines, i. a high amount of magnetic field strength can be provided. This can be advantageous if the sample is to be arranged in a sharply defined surface area.

Arranging the magnetic device relative to the sample carrier may include placing the tip relative to the predetermined region. Since a high magnetic force flux is provided in the area of the tip, the position of the tip can be used to determine the strength and position of the local extremum in the surface area.

The magnet device may comprise a conically shaped element, in particular wherein the conically shaped element comprises the tip. In particular, an iron core of an electromagnet may comprise a tip and / or correspond to a conically shaped element.

The conically shaped element may be connected to a permanent magnet or an electromagnet, be a permanent magnet, or be partially disposed inside a coil of an electrically conductive material, in particular wherein the coil is part of an electromagnet. The use of an electromagnet may be advantageous since the magnetic field, in particular the magnitude of the magnetic field strength, can be varied in this case. In particular, an electromagnet can be switched off and on. This may be particularly advantageous in the case of automation of the method.

The conically shaped element may have at the base a diameter which corresponds to the maximum spatial extent of a permanent magnet. In particular, the conical shaped element may have at the base a diameter which corresponds to the diameter of a cylindrical permanent magnet or a cylindrical iron core of an electromagnet. Thereby, an optimal connection between the conically shaped element and the permanent magnet or electromagnet can be achieved. The opening angle of the conically shaped element can be between 30 ° and 90 °, in particular 60 °.

The sample carrier may comprise an observation area, wherein the observation area is designed such that a sample arranged in the observation area can be observed by means of an optical device, for example a microscope. In particular, the desired surface area may correspond to an observation area of the sample carrier or an observation area may comprise the desired surface area. The sample carrier may comprise a bottom plate, wherein the sample carrier in operation rests on the bottom plate and wherein the magnetic device is arranged so that it is arranged in operation under the bottom plate, in particular wherein the tip of the magnetic device is disposed directly under the bottom plate.

The bottom plate may include the desired surface area. In this case, the sample can be positioned in a desired surface area of the bottom plate.

The desired magnetic field arrangement can be designed in such a way that a magnetic force acts on the introduced sample, in particular on the particles connected to the sample, so that the sample can be moved in the desired magnetic field arrangement by a magnetic force.

In particular, the magnetic force may be greater than a frictional force between the sample and a surface of the sample carrier. A liquid may be disposed in the sample carrier and, if the sample is in the liquid, the magnetic force may be greater than a viscous frictional force between the sample and the liquid. In this way, a sample can be accelerated by the magnetic force. In particular, the sample can align in the desired magnetic field arrangement and be moved along the magnetic field lines. The magnetic force can be smaller than the force with which the sample and the at least one magnetic particle are connected to each other. As a result, the sample can be moved by force transmission with the particle.

The step of placing the sample may include moving the sample carrier. In particular, the sample carrier may be moved such that, when the sample is in contact with a surface of the sample carrier, the sample is released from the surface. This may be advantageous when the magnetic force is less than a frictional force between the sample and a surface of the sample carrier. The movement of the sample carrier may include a periodic or aperiodic movement, such as jarring or pivoting of the sample carrier or vibrations by ultrasound.

• Befüllen des Hohlraums mit einer ersten Flüssigkeit,
• Einbringen einer zweiten Flüssigkeit in das Durchgangsloch, wobei die zweite Flüssigkeit eine hydrophobe Flüssigkeit ist, und
• Einbringen der Probe in die zweite Flüssigkeit.

The invention also provides a method for positioning an organic, biological and / or medical sample in a desired surface area of a sample carrier, wherein the sample carrier comprises a cavity, wherein a through hole leads into the cavity and wherein in the operation of the sample carrier, the through hole from above in leads the cavity, comprising the steps:

• Filling the cavity with a first liquid,
• Introducing a second liquid into the through-hole, wherein the second liquid is a hydrophobic liquid, and
• Introducing the sample into the second liquid.

As a result, a sample, in particular in a difficult to reach cavity of a sample carrier, can be positioned effectively and precisely.

In particular, the sample carrier may comprise one or more of the features described above.

The second liquid may have a higher viscosity, a lower density and / or a higher hydrophobicity than the first liquid. Due to the higher viscosity can be achieved that the pulse of the introduced sample is aligned parallel to the direction of gravity effect. In particular, the viscosity of the second liquid can be ten times to 10 ⁶ times the viscosity of the first liquid, in particular 10 to 1000 times or 1000 to 10 ⁶ times.

Due to the lower density of the second liquid can be achieved that the second liquid floats on the first liquid, and thereby remains arranged in the through hole. In particular, this can reduce or avoid contact instabilities at the boundary layer between the first and the second liquid, for example Rayleigh-Taylor instabilities. For example, the density of the second liquid may be between 70% and 95% of the density of the first liquid. Alternatively or additionally, an arrangement of the second liquid in the through hole can be achieved by capillary forces.

Due to the greater hydrophobicity of the second liquid, mixing of the first liquid and the second liquid can be avoided.

The first and / or second liquids may be chosen so that they do not have a toxic effect on the sample. This may be particularly advantageous when the sample is a living biological cell.

In particular, the first liquid may comprise water and / or the second liquid may comprise an oil, in particular a mineral oil and / or a silicone oil.

The sample may be introduced in the form of a suspension in the second liquid, in particular wherein the suspension comprises a third liquid, wherein the third liquid is a has stronger hydrophilicity than the second liquid. As a result, it can be avoided that the suspension mixes with the second liquid.

The second liquid may be a two-component liquid, wherein the second liquid is solidifiable after the step of introducing the sample by cross-linking or polymerizing. This allows the sample chamber to be closed. In particular, contamination of the first liquid from the outside and / or evaporation of the first liquid can thereby be avoided or reduced.

The through hole may be formed so as to taper toward the cavity. For example, the taper can be conical. By reducing the cross-sectional area of the through-hole to the cavity, a more accurate positioning of the sample is possible.

After filling with the first liquid, the first liquid may be arranged in the sample carrier such that no liquid is inside the through-hole. The second liquid may be disposed completely in the through hole after insertion. In particular, the second liquid can be introduced into the through hole such that the second liquid does not protrude beyond the outer opening of the through hole. As a result, a safe introduction of the sample into the second liquid can be achieved.

The invention also provides a positioning system for positioning an organic, biological and / or medical sample in a desired surface area of a sample carrier, comprising a magnetic device, a sample carrier holder, and a device for positioning the sample carrier relative to the magnetic device. The positioning system can be used in particular in a method described above. With the help of such a positioning system, the placement of the sample can be done precisely.

The sample carrier and / or the magnetic device may comprise one or more of the features described above.

In particular, the magnetic device may comprise a permanent magnet and / or an electromagnet.

The magnetic device may comprise a conically shaped, in particular magnetic or ferromagnetic element, in particular wherein the conically shaped element comprises a tip. In the area of the tip, a high magnetic flux of force can be provided.

The positioning system may also include means for automatically moving the magnetic device relative to the sample carrier. As a result, an at least partial automation of the positioning of the sample can be achieved. In particular, the means for automatically moving may be used to position the magnetic device relative to the sample carrier such that a desired magnetic field arrangement is provided in a predetermined region of the sample carrier. The device for automatically moving the magnetic device can be used for arranging the sample in a desired surface area of the sample carrier with the aid of the magnetic device, in particular wherein the arrangement of the sample comprises moving the magnetic device relative to the sample carrier. In particular, a more precise positioning of the sample can also be achieved by the device for automatic movement than when the method steps are carried out manually.

The positioning system may include means for automatically moving the sample carrier holder, whereby the sample carrier holder may be moved therewith such that when a sample is in contact with a surface of the sample carrier, the sample may detach from the surface. This may be particularly advantageous when arranging the sample comprises aligning the sample in the desired magnetic field order, the magnetic force being less than a frictional force between the sample and a surface of the sample carrier. The device for automated movement of the sample carrier holder may in particular comprise an ultrasonic element and / or a pivoting element. The ultrasound element can set the sample carrier holder, in particular with the sample carrier fixed therein, in vibration.

The positioning system may further comprise a pipetting device for, in particular automated, filling a sample carrier fixed in the sample carrier holder, wherein the pipetting device may comprise one or more pipettes. With the help of the pipetting device, the introduction of the sample into the sample carrier can be automated and thus made more efficient and precise.

The invention further provides a sample carrier comprising a structural element, wherein the structural element is designed such that an organic, biological and / or medical sample introduced into the sample carrier can be arranged in a desired partial area, in particular in a desired surface area, of the sample carrier. Such a structured sample carrier allows positioning of the sample in a desired surface area of the sample carrier. Such a sample carrier may be used in particular in any of the methods described above.

In particular, the sample carrier may comprise one or more of the features described above.

The sample carrier may comprise a predetermined surface area, wherein the predetermined surface area comprises the structural element, and wherein the structural element is configured such that the introduced sample is arranged in a desired portion of the predetermined surface area, in particular in the desired surface area.

The structural element may be designed in the form of an elevation and / or a depression. In particular, the structural element may be in the form of a dome, a pyramid, a groove and / or a depression.

The desired subregion can adjoin the structural element or completely surround the structural element.

In particular, the structural element may comprise the desired partial area. This may be the case, for example, if the structural element is designed in the form of a groove or a depression or comprises a groove and / or a depression.

The structural element may comprise a curved surface area or an inclined plane, in particular, so that the introduced sample can be guided along the curved surface area or along the inclined plane into the desired partial area. In particular, when the sample is a biological cell, in particular a living biological cell, it can not grow or only with difficulty on an inclined plane or a curved surface area. In particular, the sample, which is arranged after insertion in the curved surface area or the inclined plane of the structural element, can be guided by moving the sample carrier into a desired partial area.

The sample carrier may comprise a bottom plate, wherein the sample carrier rests on the bottom plate during operation, and wherein the bottom plate comprises the predetermined surface area.

In particular, the desired partial area may be partially or completely flat. In this case, the sample can stably adhere or be stably positioned in the planar region of the desired portion.

The sample carrier may comprise a bottom plate and a cover plate, wherein the cover plate and / or the bottom plate are liquid-tightly interconnected such that a cavity is formed and wherein the structural element comprises a through hole through the bottom plate or cover plate, wherein the through hole is arranged such that the sample can be placed in a desired portion of the cavity.

In this case, the structural element can be designed such that the sample can be fixed by capillary forces in the desired portion of the cavity.

The sample carrier may comprise a plastic, in particular COC (cycloolefin copolymer), COP (cycloolefin polymer), PS (polystyrene), PC (polycarbonate) or PMMA (polymethyl methacrylate). The sample carrier may be formed as an injection molded part. The bottom plate may comprise a plastic and / or glass. The bottom plate can be thin, for example between 1 .mu.m and 300 .mu.m. This allows high-resolution microscopy through the bottom plate.

• Bereitstellen eines oben beschriebenen Probenträgers,
• Einbringen der Probe in den Probenträger, und
• Bewegen des Probenträgers, so dass die Probe in dem gewünschten Oberflächenbereich des Probenträgers angeordnet wird.

The invention also provides a method for positioning an organic, biological and / or medical sample in a desired surface area of the sample carrier comprising the steps of:

• Providing a sample carrier as described above,
• Introducing the sample into the sample carrier, and
• Moving the sample carrier so that the sample is placed in the desired surface area of the sample carrier.

By moving the sample carrier, the sample can be arranged in the desired surface area. In particular, when placed in a curved portion or an inclined plane of the structural member, the sample may be directed by moving to the desired surface area.

• Bereitstellen eines Probenträgers, wobei der Probenträger eine Bodenplatte und eine Deckplatte umfasst, wobei die Deckplatte und/oder die Bodenplatte derart flüssigkeitsdicht miteinander verbunden sind, dass ein Hohlraum gebildet wird und wobei das Strukturelement ein Durchgangsloch durch die Bodenplatte oder Deckplatte umfasst, wobei das Durchgangsloch derart angeordnet ist, dass die Probe in einem gewünschten Teilbereich des Hohlraums angeordnet werden kann, und
• Einbringen der Probe in Form einer Suspension in den Probenträger.

The invention also provides a method for positioning an organic, biological and / or medical sample in a desired surface area of a slide, comprising the steps of:

• Providing a sample carrier, wherein the sample carrier comprises a bottom plate and a cover plate, wherein the cover plate and / or the bottom plate are connected to each other in such a liquid-tight manner that a cavity is formed and wherein the structural element is a through hole through the bottom plate or cover plate, wherein the through hole is arranged such that the sample can be arranged in a desired portion of the cavity, and
• Introducing the sample in the form of a suspension in the sample carrier.

In particular, the sample carrier may comprise one or more of the features described above.

The structural element can be designed such that the sample can be held by capillary forces in the desired portion of the cavity.

Before the step of introducing the sample, a gel can be introduced into the desired portion of the cavity. In particular, a Collagen1 gel, an agarose gel or a Matrigel can be used.

After the introduction of the sample, the through-hole, in particular with an optically transparent material, can be closed.

Figur 1

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement in Form einer Kuppe;

Figur 2

zeigt ein Beispiel für einen Probenträger und ein Strukturelement in Form einer Erhöhung;

Figur 3

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement in Form einer Erhöhung und eingebrachte Proben;

Figur 4

zeigt ein Beispiel für einen Probenträger und ein Strukturelement in Form einer Erhöhung;

Figur 5

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement in Form einer Erhöhung und eingebrachte Proben;

Figur 6

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement in Form einer Erhöhung und eingebrachte Proben;

Figur 7

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement in Form einer Erhöhung und eingebrachte Proben;

Figur 8

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement in Form einer Erhöhung und eingebrachte Proben;

Figur 9

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement in Form einer Erhöhung und eine optische Vorrichtung;

Figur 10

zeigt ein Beispiel für einen Teil eines Probenträgers mit Strukturelementen in Form von Erhöhungen;

Figur 11

zeigt ein Beispiel für einen Probenträger und ein Strukturelement umfassend ein Durchgangsloch durch eine Deckplatte;

Figur 12

zeigt ein Beispiel für einen Probenträger und ein Strukturelement umfassend ein Durchgangsloch durch eine Deckplatte;

Figur 13

zeigt ein Beispiel für einen Probenträger und ein Strukturelement umfassend ein Durchgangsloch durch eine Deckplatte;

Figur 14

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement umfassend ein Durchgangsloch durch eine Deckplatte und einem Gel in einem Teilbereich der Probenträgers;

Figur 15

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement umfassend ein Durchgangsloch durch eine Deckplatte und eingebrachte Proben;

Figur 16

zeigt ein Beispiel für einen Probenträger mit einem Strukturelement umfassend ein Durchgangsloch durch eine Deckplatte und einem Gel in einem Teilbereich der Probenträgers;

Figur 17

zeigt ein Beispiel für einen Teil eines Probenträgers, eingebrachte Proben und eine Magnetvorrichtung;

Figur 18

zeigt ein Beispiel für einen Teil eines Probenträgers, eingebrachte Proben und eine Magnetvorrichtung;

Figur 19

zeigt ein Beispiel für einen Teil eines Probenträgers, eingebrachte Proben und eine Magnetvorrichtung;

Figur 20

zeigt ein Beispiel für einen Teil eines Probenträgers, eingebrachte Proben und eine Magnetvorrichtung;

Figur 21

zeigt ein Beispiel für einen Teil eines Probenträgers, eingebrachte Proben und eine Magnetvorrichtung; und

Figur 22

zeigt ein Beispiel für einen Teil eines Probenträgers, eingebrachte Proben und eine Magnetvorrichtung;

Die organische, biologische und/oder medizinische Probe kann eine biologische Zelle sein. Insbesondere können eine Vielzahl von Zellen positioniert werden. Dadurch kann eine gewünschte Zellverteilung in einem gewünschten Oberflächenbereich eines Probenträgers bereitgestellt werden.Further features and advantages are explained below with reference to the exemplary figures.

FIG. 1

shows an example of a sample carrier with a structural element in the form of a dome;

FIG. 2

shows an example of a sample carrier and a structural element in the form of an increase;

FIG. 3

shows an example of a sample carrier with a structural element in the form of a raised and introduced samples;

FIG. 4

shows an example of a sample carrier and a structural element in the form of an increase;

FIG. 5

shows an example of a sample carrier with a structural element in the form of a raised and introduced samples;

FIG. 6

shows an example of a sample carrier with a structural element in the form of a raised and introduced samples;

FIG. 7

shows an example of a sample carrier with a structural element in the form of a raised and introduced samples;

FIG. 8

shows an example of a sample carrier with a structural element in the form of a raised and introduced samples;

FIG. 9

shows an example of a sample carrier having a structural member in the form of an elevation and an optical device;

FIG. 10

shows an example of a part of a sample carrier with structural elements in the form of elevations;

FIG. 11

shows an example of a sample carrier and a structural element comprising a through hole through a cover plate;

FIG. 12

shows an example of a sample carrier and a structural element comprising a through hole through a cover plate;

FIG. 13

shows an example of a sample carrier and a structural element comprising a through hole through a cover plate;

FIG. 14

shows an example of a sample carrier with a structural element comprising a through hole through a cover plate and a gel in a portion of the sample carrier;

FIG. 15

shows an example of a sample carrier with a structural element comprising a through hole through a cover plate and introduced samples;

FIG. 16

shows an example of a sample carrier with a structural element comprising a through hole through a cover plate and a gel in a portion of the sample carrier;

FIG. 17

shows an example of a part of a sample carrier, introduced samples and a magnetic device;

FIG. 18

shows an example of a part of a sample carrier, introduced samples and a magnetic device;

FIG. 19

shows an example of a part of a sample carrier, introduced samples and a magnetic device;

FIG. 20

shows an example of a part of a sample carrier, introduced samples and a magnetic device;

FIG. 21

shows an example of a part of a sample carrier, introduced samples and a magnetic device; and

FIG. 22

shows an example of a part of a sample carrier, introduced samples and a magnetic device;

The organic, biological and / or medical sample may be a biological cell. In particular, a plurality of cells can be positioned. Thereby, a desired cell distribution in a desired surface area of a sample carrier can be provided.

In general, when a sample carrier or culture vessel is filled, a random cell distribution occurs. In the case of simple bowls or pots, the cell distribution often depends on the type of filling, i. For example, how quickly the cell suspension is pipetted in and how the vessels are moved immediately after filling. In microfluidic cell culture vessels, cell distribution often depends on the geometry of the structures that receive the cell suspension.

In particular, experiments with biological cells often requires accurate positioning of the cells. As a result, the local cell density can be defined in order to be able to compare results from different experiments sufficiently well, to carry out more economically and / or to facilitate the evaluation or to enable an automation of the evaluation. For example, in a microscopic assay, it is not always necessary for cells to colonize the entire surface area of a sample carrier, but it is sufficient for cells to be arranged only in the optically accessible area or in a part thereof. This can save you rare or expensive cell material. In certain cases, it may be advantageous if cells adhere only in certain places and not the entire observation area is occupied. In this case, fewer cells consume the available medium or gas. It is thus also possible to cultivate cells under static conditions in extremely flat or small structures.

For example, the migration of adherent cells can be measured by observing the temporal evolution of the shape of an initial circular array of cells in a suitable gradient of concentration of a chemical. Remains the Form homogeneously circular over time, the cells show no directional movement, however, if the shape expands more in the direction of the gradient as in the direction perpendicular to it, a directed movement is close.

A sample carrier may comprise a plastic, in particular COC, COP, PS, PC or PMMA. An example of a sample carrier is in DE 101 48 210 described. The sample carrier may correspond to an injection molded part or comprise an injection molded part. The sample carrier may comprise a bottom plate and a cover plate. By connecting the bottom plate with the cover plate, a cavity or an upwardly open area can be formed. In the cavity may lead an opening, in particular wherein the opening for filling or emptying of the cavity, for example with the sample, can be used. The bottom plate can be connected, for example by means of fusion or gluing with the cover plate. In particular, glass can be attached by gluing. Adhesives used are, for example, UV-curing adhesives, adhesive tapes or other adhesives. In particular, substances which do not have a toxic effect on the sample can be used. Suitable welding technologies are in EP 1 579 982 described.

The sample carrier, in particular the bottom plate, may comprise a structural element, in particular a three-dimensional structural element. The structural element may be formed in the form of a survey and / or depression. The structural element can be used to position the sample, for example because a sample can not grow on a raised or rounded dome as it falls vertically or down an inclined plane.

Alternatively or additionally, a depression, for example in the form of a groove, can be formed, in which the sample can be positioned. As a result, for example, biological cells can be locally concentrated in a desired partial area.

FIG. 1 shows a sample carrier comprising a bottom plate 101 and a cover plate 102, which are connected to each other such that an upwardly open area is provided. An outburst on the side of the figure facing the observer serves as an illustration. Structural element 103 is in the form of an elevation, in particular in the form of a dome. The inner diameter of the sample carrier can be 7 mm. The structural element may have a diameter of 2 mm, a radius of curvature of the upper edge of 0.5 mm and a height of 1 mm. The structural element can be provided by deep drawing in the bottom plate 101, for example.

One or more samples may be introduced into the sample carrier. For example, the sample carrier can be filled with 100 μl of cell suspension, in particular wherein the concentration or number density of the cells in the suspension can be selected such that a desired surface area 104 can be exposed to 100% confluence with cells. 100% confluent means that no free space is visible between the cells. After the introduction of the cell suspension, for example after 30 seconds, the sample carrier can be alternately moved obliquely in opposite directions, so that cells which have deposited on the structural element 103 are guided into the desired surface area 104.

A sample carrier according to FIG. 1 can be used for a migration assay. To evaluate the migration behavior of biological cells, after a predetermined time, for example after 2 hours, an image of the sample carrier can be taken and the confluence of the cells evaluated. The percentage of confluency indicates the ratio of cell occupied area to the total area of the sample carrier surface area intended for the migration assay. On the basis of the measured data, it is possible to determine the time which is necessary, for example, to grow a flat upper region of the structural element 103 to 100% confluence.

This migration assay has significant advantages over known migration assays. For example, in the known scratch assay, a pipette tip is scraped into a cell-grown surface area to scrape a cell-free area and time measured by the cells until the scratch is closed again. Problems for reproducibility can arise, among other things, that the stud generally does not have a well-defined width, and that scratching can destroy or damage possible surface coatings of the sample carrier.

Alternatively, a region may be kept free of a sample by covering it with a silicone part that is either mechanically pressed onto the growth surface or self-adhered to the growth surface by a tacky layer. Experimental arrangements that use silicone parts to make cell-free regions in confluent cell cultures have the disadvantage that the silicone parts must be removed prior to the actual assay and create an additional risk of contamination. In addition, coating proteins may also adhere to the silicone, which may interfere with any protein coating of a surface area of the sample carrier. Due to the relatively high elasticity of the silicone, the accuracy of the size of the recessed area is limited.

A sample carrier as in FIG. 1 does not include any moving parts in contact with the sample. The dimensioning of the structural element 103 can be reproducible, for example, by means of a correspondingly optimized thermoforming process.

FIGS. 2 to 5 each show a cross section through a sample carrier with a structural element 203, 303, 403 or 503. The structural element 203 or 303 in FIGS. 2 and 3 is designed in the form of a pyramid. As a result, the structural element 203 or 303 comprises a plurality of inclined planes. In particular, the structural element 203 or 303 in FIG FIGS. 2 and 3 a dull pyramid, ie the tip is flattened.

FIG. 4 and FIG. 5 show a structural element 403 or 503 in the form of a dome with vertical walls.

In FIGS. 3 and 5 For example, individual samples 305 and 505 are shown, which are arranged in a desired surface area.

The sample carrier comprises in each case a bottom plate 201, 301, 401 or 501 and a cover plate 202, 302, 402 and 502, respectively.

FIGS. 6 to 8 show a cross section through a sample carrier comprising a structural element 603, 703 or 803. The samples 605, 705 and 805 are arranged in a medium 606, 706 or 806, in particular a liquid. In FIG. 6 The filling height of the liquid 606 corresponds to the height of the structural element 603. FIG. 7 shows the sample carrier FIG. 6 at a later time, with the samples 705 arranged in a desired surface area of the sample carrier. In other words, the samples 705 have dropped to the bottom of the sample carrier and adhere there. FIG. 8 shows the sample carrier from the FIGS. 6 and 7 wherein the sample carrier is filled with the medium 806 up to a predetermined filling level.

The sample carrier each comprises a bottom plate 601, 701 or 801 and a cover plate 602, 702 and 802, respectively.

FIG. 9 shows a sample carrier comprising a cavity 907, wherein the cavity 907 comprises an observation channel 908. In the observation channel 908, a structural element 903 is arranged. A sample carrier as in FIG. 9 can be used, for example, for a chemotactic experiment. For this purpose, a gradient of a chemical substance between two subregions of the cavity 907 is built up, for example by filling only a portion of the cavity 907 with this chemical substance. An optical system 909, in particular a microscope, can be used to monitor the movement of the samples 905, especially living biological cells. The focus of the viewing optics 909 can be adjusted so that only samples located at the highest point of the feature 903 are sharply imaged. For this, the structural element 903 may comprise a round or flattened tip.

FIG. 10 shows a surface region of a sample carrier, in particular a surface region of a bottom plate 1001, comprising three structural elements 1003, wherein each of the structural elements 1003 is formed as an elongated elevation. It is also possible to use structural elements in the form of elongated depressions or to combine elongate depressions with depressions, for example with depressions of different diameters. Height and width of the strip-shaped structural elements can be varied.

FIGS. 11 to 13 show a sample carrier comprising a cavity 1107, 1207 or 1307, a bottom plate 1101, 1201 and 1301, respectively, and a cover plate 1102, 1202 or 1302 connected to the bottom plate 1101, 1201 and 1301, respectively. A structural element 1103, 1203 or 1303 comprises an opening 1111 or 1211 in the cover plate 1102, 1202 or 1302. The opening 1111 or 1211 is in particular conical, in particular wherein the opening 1111 or 1211 tapers towards the bottom plate 1101, 1201 and 1301, respectively. Through the opening 1111 or 1211, a sample 1205 or 1305 can be introduced in the form of a suspension 1110 in the sample carrier (see Fig. 11 ). The amount of suspension may be such that, as in Fig. 12 is shown, the observation area 1208 is filled and a portion of the suspension 1110 is disposed in the opening 1211. A liquid outlet from the observation area 1108, 1208 or 1308 in a first or second portion of the cavity 1207 is prevented by capillary effects. The samples can sink in the region of the opening 1111 or 1211 to the bottom of the observation area 1108, 1208 or 1308 and adhere there. After adhering, the cavity 1107, 1207 or 1307 can be filled. The opening 1111 or 1211 can be sealed and sealed with an optically transparent material, for example PDMS (polydimethylsiloxanes, eg Sylguard 184, Dow Corning Corporation). A filled sample carrier with a closed opening is in Fig. 13 shown.

FIG. 14 shows a sample carrier comprising an observation area, wherein in the observation area a piece of gel 1412, for example Collagen1 gel, agarose gel or Matrigel (for example, by Becton Dickinson), is arranged. If the sample 1505 (in the form of a suspension 1410) is filled in the sample carrier, as in FIGS. 15 and 16 As they are shown, they sink to the surface of the gel, adhere there and can migrate or sink into the gel. As a result, the cells can be arranged in a spatial area above the desired surface area. In other words, for several samples, a three-dimensional distribution of the Samples in the gel can be achieved. In addition, the show FIGS. 14 to 16 a sample carrier comprising a bottom plate 1401, 1501 or 1601, a cover plate 1402, 1502 or 1602, a cavity 1407, 1507 or 1607 and a structural element 1403, 1503 or 1603. In the observation area is a piece of gel 1412, 1512 or 1612 arranged. In FIGS. 14 and 15 an opening 1411 or 1511 is shown in the structural element 1403 or 1503 in the form of a through hole through the cover plate 1402 and 1502, respectively.

For positioning a sample in a sample carrier comprising a cavity and an opening which leads into the cavity, the following method is suitable.

Above the desired surface area of the sample carrier, in particular over an observation area of the sample carrier, there may be an opening which leads from the outside into the cavity. First, the cavity can be filled with a medium, in particular wherein the medium does not enter the opening via the height of the cavity. The medium may comprise a nutrient medium for biological cells and in particular correspond to a first liquid. The opening can then be closed with a second liquid, in particular a drop of oil, for example silicone oil or mineral oil, wherein only so much is filled that the oil surface does not bulge upwards. The sample may be added to the oil in the form of a suspension. The sample sinks through the oil to the desired surface area and can adhere or grow there. The sample can be accurately positioned. In particular, multiple samples can be positioned, with the number of samples being precisely adjustable. As a result, it is possible to work with a smaller number of samples and the sample can also be positioned in hard-to-reach surface areas of the sample carrier.

In particular, experimental preparations may be made prior to introduction of the sample. For example, a concentration gradient can be established in the sample carrier before the sample is introduced into the sample carrier. The idea is to introduce samples, in particular cells, into an experimental environment only when all or a majority of the experimental parameters, for example the gradient of a chemical substance, the temperature, the gas concentration in the medium and / or the ph value, are set. Thus cells are not disturbed by the preparations of the experiment, which can happen, for example, by changes in solution, vibrations or temperature fluctuations. In this way, the cells can be in a (maximally) comparable state at the beginning of the experiment. Immediately after insertion, the cells may be in the desired gradient so that the response of the cells can be observed without delay. Also slightly or non-adherent cells, ie cells that are not on a surface of the sample carrier can be examined with this method. Examples are immune cells, for example neutrophils and other leukocytes. Since the oil largely prevents evaporation of the first liquid, it is possible in particular to work with small amounts of medium.

For example, a sample carrier comprising two reservoirs and an observation channel disposed therebetween, such as in FIG EP 1 741 487 described, filled with samples. For this purpose, the sample carrier is first filled with a neutral medium. Subsequently, a gradient of a chemical substance is built up between the reservoirs. Since this can take a certain time, in particular a few hours, it is possible by this method that the sample is introduced only in the fully established gradient. Directly above the observation channel may be an opening, which is for example conical and closed with a hydrophobic liquid. The hydrophobic liquid may be, for example, a silicone oil or a mineral oil, in particular where the oil is selected such that it does not act toxic to the sample and does not arouse or destroy the materials of the sample carrier. As a hydrophobic liquid, a two-component liquid can be used, which is introduced into the filling opening shortly before the introduction of the sample and then polymerize or otherwise cross-link and solidify. Examples of these are silicone oils mixed with crosslinkers or, for example, Sylguard 184 from Dow Corning (PDMS). Once the sample has been introduced, the observation can be carried out, for example with the aid of a microscope.

A positioning of a sample in a desired surface area of a sample carrier can be carried out by means of a magnetic force. For this purpose, the sample must have magnetic properties and be exposed to magnetic forces in a corresponding sample carrier. Biological cells usually have no magnetic properties. In order to magnetically manipulate cells as a sample, they must be "magnetized". For this example, paramagnetic particles, in particular paramagnetic nanoparticles are suitable. The particles can be connected to the sample in a variety of ways. Small particles can be phagocyted by cells, that is, taken up. Prerequisite for the inclusion is an attachment of the particles on the surface of the cell. In particular, positively charged end groups are suitable for attachment to the surface of the cell, since the cell membrane usually carries negative charges. The particles can be stored, for example, in vesicles in the cytosol. With an appropriate amount of collected particles, the external action of a magnetic field may be large enough to move a non-adherent cell in a sample carrier.

Another possibility is binding of the particles to the cell surface. Here, the magnetic particles may be larger, i. be nearly as big as the cell itself or bigger. In particular, the size of a particle may correspond to one-fiftieth of the cell size. The particles may, for example, consist of a paramagnetic material in their core and may be coated with a polymer matrix. On this polymer matrix, the particles may have a coating that can adhere to a cell surface. Examples include surface proteins such as CD molecules or activated tosyl groups. The binding of the particles to the cells may be specific or nonspecific by the choice of coating. In particular, the coating can be chosen so that it adheres to only one type of cells, that is specific. As a result, a desired type of cells can be filtered out of a multiplicity of cells.

In order to have a force acting on the sample, in particular on a cell, a magnetic field can be applied, in particular perpendicular to the potential direction of movement, for example to the growth surface of the sample carrier. For concentrating a plurality of samples in a defined, radially symmetrical surface area, for example, a field can be created whose field lines condense to the desired surface area. If one strives for a circular cell spot, the field in this area can be strongest and in concentric circles around the desired surface area the field lines can become less dense. This can be achieved, for example, with an iron cone, the tip of which is placed directly below the desired surface area.

FIGS. 17 to 20 show a part of a sample carrier, in particular an observation channel 1708, 1808, 1908 or 2008, comprising a bottom plate 1701, 1801, 1901 and 2001 and a cover plate 1702, 1802, 1902 and 2002, respectively. A cone or conically shaped element 1713, 1813 , 1913 or 2013 of a magnetic or magnetizable material is connected to a permanent magnet 1714, 1814, 1914 and 2014, respectively. The permanent magnet 1714, 1814, 1914, and 2014, for example, may be a neodymium-iron-boron (NdFeB) magnet. The amount of field strength of the permanent magnet 1714, 1814, 1914, and 2014, respectively, may be between 0.5 and 1.4 Tesla. The magnetic field is collimated to the top of the conical element 1713, 1813, 1913, and 2013, respectively, and a magnetic field line distribution is formed in which the field lines at the tip of the conical shaped element 1713, 1813, 1913, and 2013, respectively, are highly densified. The permanent magnet 1714, 1814, 1914 and 2014 may have a diameter between 1 mm and 20 mm, in particular 3 mm to 10 mm. The conically shaped element 1713, 1813, 1913, and 2013, respectively, may have a diameter at the base that corresponds to the diameter of the permanent magnet 1714, 1814, 1914, and 2014, respectively. The opening angle of the conically shaped element 1713, 1813, 1913 and 2013 may between 30 ° and 90 °, in particular 60 °. For a positioning of a sample in an observation channel 1708, 1808, 1908 or 2008, for example 1 mm wide and 70 μm high, a conically shaped element 1713, 1813, 1913 or 2013 with a diameter of the base of 4 mm is suitable. At the top, the conically shaped element 1713, 1813, 1913 and 2013 may taper to a flattened tip, wherein the flattened portion may have a diameter of 0.5 mm.

The opening angle of the conically shaped element 1713, 1813, 1913 and 2013 may be 60 °. The permanent magnet 1714, 1814, 1914 and 2014 may have a diameter and a height of 4 mm. Instead of a permanent magnet 1714, 1814, 1914 and 2014, an electromagnet can be used. This can be advantageous for automation of the method since the magnetic field of an electromagnet varies, in particular can be switched on and off.

The magnetic device can be positioned relative to the sample carrier. In particular, the position of the magnetic device can be changed parallel to the sample carrier, as in Fig. 17 indicated, or perpendicular to it, as in FIGS. 18-20 shown. For example, the desired magnetic field arrangement, in particular the strength of a local extremum of the magnitude of the magnetic field strength, can be varied by the perpendicular distance to the sample carrier. FIGS. 18 to 20 show the magnetic device at different distances to the sample carrier. As a result, the diameter of the desired surface area in which samples 1705, 1805, 1905 and 2005 are arranged can be varied.

FIGS. 21 and 22 show a magnetic device 2114 or 2214 and a part of a sample carrier, in particular an observation channel 2108 or 2208, comprising a bottom plate 2101 or 2201 and a cover plate 2102 or 2202. The magnetic device 2114 or 2214 has a tip 2115 and 2215, respectively Shape of a cuboid extension on. As in Fig. 21 indicated, the magnetic device can be positioned relative to the sample carrier. In particular, the size of the desired surface area can be determined by the perpendicular distance of the tip 2115 or 2215 from the observation channel 2108 or 2208. For example, shows Fig. 22 in that when the tips 2115 and 2215 are positioned closer to the viewing channels 2108 and 2208 respectively, the samples 2105 and 2205, respectively, are placed in a smaller surface area of the sample carrier. This can be explained by a more developed local extremum of the magnitude of the magnetic field strength of the desired magnetic field arrangement.

The samples can be introduced, for example, in a suspension into the sample carrier, wherein the number density of the samples in the suspension corresponds to the desired cell density. The suspension can be introduced with a pipette, wherein the entire liquid of the suspension can flow over the position of the magnetic field peak. Here, the cells are held in the magnetic field, but not immediately concentrated at the top. This method can be used for observation channels. In this case, the liquid is necessarily rinsed past the desired magnetic field arrangement with suitable positioning of the magnetic device.

To densify the samples in the desired surface area, the samples, before they can adhere to a surface region of the sample carrier, are caused by small shocks or vibrations in motion. The samples, in particular the cells, move in the direction of the reinforcing field lines. In other words, they can be gradually shaken to a maximum of magnetic field strength. The movement or small impacts can be achieved by vibrations on a shaker, by ultrasound or by pivoting the sample carrier.

After the sample has been positioned, the entire experimental set-up, in particular the sample-carrying sample carrier and the magnetic device, can be placed in an incubator for adhesion. This may take several hours. Only after this time, the magnetic device can be removed.

The above-described methods and / or sample carriers can be combined in any desired manner.

For example, a sample carrier can be used for chemotactic assays, where migration of cells in a gradient is to be monitored. The aim is to analyze whether cells migrate more or less in the direction of the increasing concentration of a substance. For this purpose, closable reservoirs can be connected by an observation channel, the height of the observation channel being less than 10% of the height of the reservoirs, for example 70 μm at a reservoir height of 800 μm. Through openings, cells and solutions can be filled into the reservoirs.

In the middle of the observation channel, a groove can be introduced into the bottom plate perpendicular to the connecting line of the reservoirs, the profile of the groove having a maximum height of, for example, 100 μm and a maximum width of, for example, 100 μm. The length of the groove may correspond to the width of the observation channel. First, both reservoirs can be filled with a neutral liquid. The neutral liquid can one Nutrient liquid for cells correspond. Then you put in one of the reservoirs cells that have been made magnetic, for example, by phagocytosis of magnetizable particles. Then, a permanent magnet can be placed under the cell-filled reservoir. This magnet can then be moved in the direction of the second reservoir. The cells follow the movement of the magnetic device until they get stuck in the groove. There you can let the cells adhere. Subsequently, a gradient of a chemical substance can be built up in the observation channel.

As an alternative to the groove, several round indentations with pointed or flat, horizontal bottom can be introduced. In this case, the magnetic cells may be introduced into the wells by moving the sample carrier systematically relative to the magnet. The maximum radii of the depressions may be, for example, 50 μm to 1 mm, and the maximum depth of the depressions may be approximately μm to 100 μm.

Instead of introducing the cells into depressions, it is also possible to produce a structure protruding from the base plate, for example by deep drawing or hot embossing of a plastic film. The structural element may correspond to a rectangular barrier whose longitudinal direction is perpendicular to the connecting line of the two reservoirs. The width of the barrier may correspond approximately to the maximum distance traveled by a cell during the observation period. Typical observation periods are for example 12 or 24 hours. In 12 hours, for example, human endothelial cells such as HU-VEC, on average, recover 200 μm in the direction of a well-defined gradient, or 400 μm in 24 hours. Over a length of about 200 μm to 400 μm, the migration of many cell types from mammals to chemotaxis needs to be analyzed and assessed. Therefore, a barrier width between 50 microns and 1000 microns can be selected.

The experiment can be carried out such that cells are introduced into the observation channel and removed by tilting the sample carrier from the barrier-shaped structural element. With the aid of a magnetic device, cells can be positioned in a partial area or removed from a partial area which adjoins the barrier-shaped structural element. The observation of the migration of the cells can be carried out by means of video microscopy. It can thereby be determined whether significantly more cells migrate in the direction of the increasing or decreasing concentration of the chemical substance. The barrier width can also be less than 50 microns. In particular, the structural element can not comprise a flat but, for example, only a curved region. If the cells are fluorescent, for example, by a GFP construct (Green Fluorescent Protein construct), the barrier-crossing Visualize cells by appropriate focusing when they are near the highest region of the feature.

Alternatively, for example, one can generate a single image at the end of the observation period and evaluate the cell distribution on the structural element. For this purpose, by means of image processing, strips of the width of a cell diameter can be superimposed on the region of the structural element, the strips extending in the longitudinal direction of the barrier. For each strip, the cells can be counted and the number of cells versus the distance of the strip from one end of the barrier applied. If one chooses the observation period so that the cells can run maximally to the middle of the structural element, for example, a higher cell density on the barrier side, which points in the direction of the decreasing gradient, an indication of chemotactic activity.

It is understood that the features mentioned in the embodiments described above are not limited to these specific combinations and are also possible in any other combinations. In particular, different sample carriers can be combined with different methods for positioning an organic, biological and / or medical sample.

Claims (13)

Sample carrier comprising a structural element, wherein the structural element is designed such that an introduced into the sample carrier organic, biological and / or medical sample in a desired portion, in particular in a desired surface area, the sample carrier can be arranged. Sample carrier according to claim 1, wherein the sample carrier comprises a predetermined surface area, wherein the predetermined surface area comprises the structural element, and wherein the structural element is formed such that the introduced sample in a desired portion of the predetermined surface area, in particular in the desired surface area, is arranged. Sample carrier according to claim 1 or 2, wherein the structural element is designed in the form of an elevation and / or a depression, in particular in the form of a dome, a pyramid, a groove and / or a depression. Sample carrier according to claim 2 or 3, wherein the desired subregion adjoins the structural element or completely surrounds the structural element, in particular wherein the structural element comprises the desired subregion, in particular if the structural element is in the form of a groove or a depression or a groove and / or a Valley includes. Sample carrier according to one of the preceding claims, wherein the structural element comprises a curved surface area or an inclined plane, in particular, so that the introduced sample can be guided along the curved surface area or along the inclined plane into the desired partial area. Sample carrier according to one of the preceding claims, wherein the sample carrier comprises a bottom plate, wherein the sample carrier rests in operation on the bottom plate, and wherein the bottom plate comprises the predetermined surface area. Sample carrier according to one of the preceding claims, wherein the desired partial area is partially or completely flat. Sample carrier according to one of the preceding claims, wherein the sample carrier comprises a bottom plate and a cover plate, wherein the cover plate and the bottom plate are connected in such a liquid-tight manner that a cavity is formed and wherein the structural member comprises a through-hole through the bottom plate or cover plate, wherein the through-hole is arranged such that the sample is locatable in a desired portion of the cavity. Sample carrier according to one of the preceding claims, wherein the inner diameter of the sample carrier is 7 mm and the structural element has a diameter of 2 mm, a radius of curvature of the upper edge of 0.5 mm and a height of 1 mm. Sample carrier according to one of the preceding claims, wherein the structural element is produced by deep drawing or hot embossing. Sample carrier according to one of the preceding claims, wherein the sample carrier comprises a bottom plate having the structural element, in particular wherein the bottom plate has a thickness between 1 .mu.m and 300 .mu.m and / or in particular wherein the structural element is produced by deep drawing or hot embossing in the bottom plate , Method for positioning an organic, biological and / or medical sample in a desired surface area of a sample carrier, comprising the steps: Provision of a sample carrier according to one of the preceding claims, Introducing the sample into the sample carrier, and Moving the sample carrier so that the sample is placed in the desired surface area of the sample carrier. Method, in particular according to claim 12, for positioning an organic, biological and / or medical sample in a desired surface area of a sample carrier, wherein the sample carrier comprises a cavity, wherein a through hole leads into the cavity and wherein in the operation of the sample carrier, the through hole from above into leads the cavity, comprising the steps: Filling the cavity with a first liquid; Introducing a second liquid into the through-hole, the second liquid being a hydrophobic liquid; and Introducing the sample into the second liquid.

Images