CN102023379B - Three-dimensional optical tweezers system - Google Patents
Three-dimensional optical tweezers system Download PDFInfo
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- CN102023379B CN102023379B CN2009100930300A CN200910093030A CN102023379B CN 102023379 B CN102023379 B CN 102023379B CN 2009100930300 A CN2009100930300 A CN 2009100930300A CN 200910093030 A CN200910093030 A CN 200910093030A CN 102023379 B CN102023379 B CN 102023379B
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Abstract
The invention provides a three-dimensional optical tweezers system. The system comprises a laser output unit, a spatial light modulator, a laser converging unit, a sample cell and a main observation unit, wherein the laser converging unit comprises a converging unit lens assembly and a front objective lens; the converging unit lens assembly is used for expanding each laser beam output by the spatial light modulator; the front objective lens is used for converging each expanded laser beam into the sample cell; the main observation unit comprises a visible light source, a rear objective lens and a main observation unit, namely a charge coupled device (CCD); the optical axes of the visible light source, the front objective lens, the rear objective lens and the main observation unit CCD are collinear to form a main optical axis; the sample cell is positioned between the front objective lens and the rear objective lens and is adjacent to both the front objective lens and the rear objective lens; both the front objective lens and the rear objective lens can move along the main optical axis; and the visible light source is positioned on the main optical axis of the front objective lens. By the system, a three-dimensionally distributed light trap can be observed under the condition that one surface of the sample cell is non-transparent. The system is particularly suitable for observation and operation of a biological living sample.
Description
Technical field
The present invention relates to optical technical field, specifically, the present invention relates to a kind of three-dimensional light tweezers system.
Background technology
Optical optical tweezers system is a kind of optical system of utilizing ligh trap to move molecule.Wherein, ligh trap is by high-NA objective laser to be assembled by force to form, and it is more severe that laser is assembled, and the gradient force of ligh trap is big more, helps fettering molecule more.The ligh trap that forms with the parallel laser of the parallel incident of objective lens optical axis is at the center of object lens front focal plane; There is the parallel laser of certain angle incident to form ligh trap with objective lens optical axis in the off-centered place of object lens front focal plane; Have the ligh trap and the object lens front focal plane of the incident laser formation of certain angle of divergence that certain distance is arranged, this distance is relevant with the degree of divergence of laser.
The laser that comes out from laser instrument incides behind beam-expanding collimation the high-NA objective, forms the monochromatic light trap.In traditional optical optical tweezers system, beam of laser forms a ligh trap, if will form two or more ligh traps, then need beam of laser be divided into the orthogonal laser of two bundle polarizations; Be converged to two or more ligh traps by same object lens after the laser alignment that also can come out by two or more laser instruments.These methods all require to become a branch of back to get into object lens the beam convergence that separates, and this brings certain degree of difficulty for building of light path, and the ligh trap quantity that forms is very limited.If will form movably ligh trap, then need use acoustooptic deflector or other rotating mirror apparatus that laser is deflected, thereby change the focus point position of laser at the object lens front focal plane.In list of references " Carlos Bustamante; Yann R.Chemla; Jeffrey R.Moffitt; High-resolution Dual Trap Optical Tweezers with Differential Detection, " Single-molecule techniques:a laboratory manual " the 297th page, Cold Spring Harbor Laboratory Press ", told about in detail and how to have realized moving of two optical optical tweezers systems and ligh trap.
List of references " Jennifer E.Curtis; Brian A.Koss; David G.Grier, Dynamic holographic optical tweezers, Optics Communications 207 (2002) 169-175 " discloses a kind of optical optical tweezers system that adopts spatial light modulator.Spatial light modulator is a kind of device that the space distribution of light wave is modulated, and it carries out the conversion or the modulation of room and time to the Two dimensional Distribution of some characteristic of light wave such as phase place, amplitude, intensity, frequency, polarization state etc. under the control of control signal.In the optical optical tweezers system; Adopt spatial light modulator through the wavefront of laser being modulated the distributed in three dimensions that can realize ligh trap; The position that is each light tweezer can be at three dimensions synchronously or independent variation, and the quantity of light tweezer, position and light intensity are distributed and can be controlled through spatial light modulator.But with reference to Jennifer literary composition, can find out, in this article disclosed three-dimensional light tweezer be object lens through being arranged in the sample cell below with beam convergence to sample cell, simultaneously, also the sample in the sample cell is carried out to picture through these object lens.This design makes CCD can only know the image of the specific focal plane that observes object lens converging parallel light beam and form, and is difficult to the ligh trap of distributed in three dimensions is carried out overall situation observation.And because the disclosed optical optical tweezers system of Jennifer literary composition adopts the transmission-type imaging, the necessary two sides of its sample cell is all transparent, so the range of application of this optical optical tweezers system is restricted.
Summary of the invention
The purpose of this invention is to provide a kind of three-dimensional light tweezers system that can observe the distributed in three dimensions of ligh trap easily,, can realize better ligh trap is controlled through observation to the ligh trap distributed in three dimensions.
For realizing the foregoing invention purpose, the invention provides a kind of three-dimensional light tweezers system, comprising: laser output unit, spatial light modulator, laser convergence unit, sample cell and subjective measurement unit;
Said spatial light modulator is used for the single beam laser of said laser output unit output is divided into multiple laser;
Said laser convergence unit comprises convergence unit lens combination and preceding object lens, and said convergence unit lens combination is used for each Shu Jiguang of spatial light modulator output is expanded bundle, said before object lens be used for respectively restrainting laser convergence to said sample cell behind the bundle with expanding;
Said subjective measurement unit comprises visible light source, back object lens and subjective measurement unit CCD,
The optical axis conllinear of said visible light source, preceding object lens, back object lens and subjective measurement unit CCD forms primary optical axis; Said sample cell between preceding object lens and back object lens and with said before object lens all adjacent with back object lens; Object lens all can move along said primary optical axis with the back object lens before said, and said visible light source is positioned on the primary optical axis of preceding object lens one side.
In the technique scheme; Said laser convergence unit also comprises convergence unit CCD and the semi-transparent semi-reflecting level crossing of convergence unit; The semi-transparent semi-reflecting level crossing of said convergence unit is used for the light path of assembling unit CCD is coupled into said primary optical axis between visible light source and preceding object lens.
In the technique scheme, said laser output unit is used to export the laser of infrared band, and said convergence unit also comprises the anti-infrared visible level crossing of convergence unit, is used for the light path of said convergence unit lens combination is coupled into said primary optical axis.
In the technique scheme; Said laser output unit is used to export the laser of infrared band; Said three-dimensional light tweezers system also comprises small location measurement unit; Said small location measurement unit comprises the anti-infrared visible level crossing of the location measurement unit between back object lens and subjective measurement unit CCD, and the light path of said small location measurement unit is coupled into said primary optical axis through the anti-infrared visible level crossing of said location measurement unit.
In the technique scheme, said laser output unit is used for the laser that output wavelength is 1064nm.
In the technique scheme, said primary optical axis is parallel to surface level.
Compared with prior art, the present invention has following technique effect:
1, the present invention can observe the ligh trap that is positioned at the sample cell different depth through moving the back object lens, thereby has realized the observation to the ligh trap of distributed in three dimensions.
2, the scope of application of the present invention is wider, and under the opaque situation of sample cell one side, the present invention still can observe through convergence unit CCD.
3, the present invention is particularly suitable for the biological living sample is observed and operated.
Description of drawings
Below, specify embodiments of the invention in conjunction with accompanying drawing, wherein:
Fig. 1 shows the light path synoptic diagram of one embodiment of the invention;
Fig. 2 shows the phase diagram of one embodiment of the invention spatial light modulator and the optical trap array that is observed; Two phase diagrams when wherein (a), (c) part be the spatial light modulator modulated laser, (b), (d) part is the image that is respectively the bead that (a), the pairing optical trap array of (c) part phase diagram catch.
The drawing explanation
1 first lens, 2 half-wave plates, 3 second lens
4 first catoptrons, 5 spatial light modulators, 6 second catoptrons
The anti-infrared visible level crossing of 7 the 3rd lens 8 the 4th lens, 9 convergence units
10 the 5th lens 11LED lamps, 12 visible light wave range semi-transparent semi-reflecting lens
Anti-infrared visible level crossing 18 the 8th lens in 16 the 6th lens, 17 displacement measurement unit
19 observing unit CCD, 20 observing unit monitors 21 the 7th lens
22 convergence unit CCD, 23 convergence unit monitors, 24 Position-Sensitive Detectors
25 laser instruments
Embodiment
Fig. 1 shows the light path synoptic diagram of the multiple light forceps system of one embodiment of the invention.The multiple light forceps system of this embodiment comprises laser instrument 25; First lens 1; Half-wave plate 2; Second lens 3; First catoptron 4; Spatial light modulator 5; Second catoptron 6; The 3rd lens 7; The 4th lens 8; The anti-infrared visible level crossing 9 of convergence unit; The 5th lens 10; LED lamp 11; Visible light wave range semi-transparent semi-reflecting lens 12; Preceding object lens 13; Sample cell 14; Back object lens 15; The 6th lens 16; The anti-infrared visible level crossing 17 in displacement measurement unit; The 8th lens 18; Observing unit CCD19; Observing unit monitor 20; The 7th lens 21; Convergence unit CCD22; Convergence unit monitor 23 and Position-Sensitive Detector 24.
Wherein, laser instrument is a 1064nm high power CW laser instrument, and peak power is 4W, and when this had guaranteed to form many ligh traps, each ligh trap can both be assigned to enough laser powers.This helps the stable granule of catching of ligh trap.If laser power is too low, again more for a long time, the laser power that each ligh trap is assigned to is less, causes ligh trap power less at the ligh trap number that forms, and this is unfavorable for catching molecule very much.In the present embodiment, laser instrument adopts the Compass 1064-4000M laser instrument of Coherent company.
The laser that laser instrument comes out is reflexed in the spatial light modulator 5 by first catoptron 4 and second catoptron 6 behind first lens 1 and second lens, 3 expansion bundles.After expanding bundle, it is big that laser beam diameter becomes, and the angle of divergence diminishes, in perpendicular to the xsect of optical axis direction light distribution more even, more approach directional light.The expansion bundle helps object lens laser is converged to littler hot spot at the object lens front focal plane, and is also bigger in the intensity gradient at ligh trap place.In the present embodiment, can also increase a half-wave plate at first lens 1 and 3 on second lens, this half-wave plate is used for the polarization direction of laser is rotated to the direction that spatial light modulator is the most effectively modulated.
The phase diagram of spatial light modulator is in advance good according to the ligh trap distribution requirements; The phase place of using this phase diagram to advance to modulate spatial light modulator; Multiple laser after then spatial light modulator being modulated imports the 3rd lens 7 and the 4th lens 8 expand bundle; Each Shu Jiguang that will expand again after restrainting incides preceding object lens 13 (in the present embodiment, most of light beam of object lens 13 is a parallel beam before inciding, and these light beam incident directions are parallel with the optical axis of preceding object lens 13); Said the 3rd lens 7 and the 4th lens 8 are except expanding bundle, and another effect is on the back focal plane of object lens 13 before spatial light modulator 5 is imaged onto.Laser forms ligh trap after preceding object lens the last 13 assembled like this.
In the present embodiment, adopt the spatial light modulator of HOLOEYE company, model is: PLUTO-0,7 " HDTV LCOS Phase Only Kit Modulator for phase displays VIS, NIR&TELCO HES:6010xxx.This spatial light modulator is pure phase-type reflective spatial light modulator, and the incident angle when laser incides spatial light modulator requires to be not more than 5 °.For making full use of each pixel of spatial light modulator, the laser beam diameter that is input to spatial light modulator is generally greater than the size of spatial light modulator photosurface.The focal length of the 3rd lens 7 and the 4th lens 8 is decided according to the input aperture of preceding object lens 13 and the photosurface size of spatial light modulator 5.Generally, the entrance pupil of object lens before laser will be full of.
In the present embodiment, its sensitive wave length of spatial light modulator is near infrared (850-1100nm), and pixel is 1920*1080, and each pixel is of a size of 8 μ m, and the pixel fill factor, curve factor is 0.87, largest frames frequency 60Hz.Laser is converged to ligh trap by object lens, can regard as carrying out the process of Fourier transform before the laser wave.The wavefront of modulation incident laser just can reach the purpose of modulating ligh trap.If the amplitude of modulated laser wavefront can lose a part of laser power, this is unfavorable to ligh trap efficient, so present embodiment is taked the only way of modulated laser Wave-front phase.Therefore, as long as spatial light modulator just can realize the modulation to ligh trap according to the phase place of certain phase diagram modulated laser wavefront.The computing method of contraposition phasor have detailed introduction in list of references " Eric R.Dufresne, Gabriel C.Spalding, Matthew T.Dearing; and Steven A.Sheets, David G.Grier, Computer-generated holographic optical tweezer arrays; REVIEW OF SCIENTIFIC INSTRUMENTS VOLUME 72; NUMBER 3, and MARCH 2001 " and list of references " Jennifer E.Curtis, Brian A.Koss; David G.Grier; Dynamic holographic optical tweezers, Optics Communications 207 (2002) 169-175 ", therefore repeat no more here.
In the present embodiment, it is the zeiss object lens of 420792-9900-000 that preceding object lens 13 adopt model, and these object lens are N.A=1.4, the oil immersion objective of 100X.
It is the leica object lens of HCX PL FL L 63x/0.70CORRPH2 that back object lens 15 adopt model.For reaching desirable imaging effect, preceding object lens 13, back object lens 15 will be strictly with incide before the laser coaxial of object lens 13, simultaneously, the visible light of LED lamp 11 outgoing, the 5th lens 10, the 6th lens 16 also will with incide before the laser coaxial of object lens 13.In addition, preceding object lens 13, sample cell 14, back object lens 15 all are installed on the electronic accurate translation stage, can guarantee the degree of accuracy and the degree of stability of system like this.
The back focal plane that the 8th lens 18 are incited somebody to action back object lens 15 is imaged onto Position-Sensitive Detector 24, and (English name of Position-Sensitive Detector is position-sensitive detector; Be abbreviated as PSD) on, reacted the conoscope image of laser after the ligh trap place is by the granule scattering on the back focal plane of back object lens 15.It is very sensitive that the conoscope image of laser departs from the ligh trap center displacement to bead, therefore can very accurately measure the micro-displacement of bead in ligh trap.
Present embodiment has transmission-type and the reflective pair of imaging system and micro-displacement measuring system.
With reference to the accompanying drawings 1, further this operation of overlapping three-dimensional multiple light forceps system is elaborated:
Regulate back object lens 15 in the position of optical axis direction, make the sharply defined image at the bottom of observing unit CCD19 presents sample cell.LED lamp 11, back object lens 15, the 6th lens 16 and observing unit CCD19 have constituted the transmission-type imaging system.Continue to regulate back object lens 15 in the position of optical axis direction, can in observing unit CCD19, see the picture of different depth in the sample cell continuously.If ligh trap is three-dimensional ligh trap, can see that any one ligh trap catches the situation of molecules such as bead or cell through the position of regulating back object lens 15 so.
As shown in Figure 2, the phase diagram when (a) part is a spatial light modulator modulation.The effect of this phase diagram is to generate a circle to be distributed in eight ligh traps on the ring.(b) behind part phase diagram for employing (a) part in the experiment, the sample drawing that observing unit CCD19 collects, at this moment, laser power is 500mW, bead is diameter 3 μ m polystyrene spheres.Can find out that from (b) part of Fig. 2 one has 9 ligh traps has captured bead, the bead at its centre circle ring center is caught by the zero level hot spot.If the ligh trap of not wanting the zero level hot spot to form can be put into a light barrier between the 3rd lens 7 and the 4th lens 8, keep off the zero level hot spot.The way of light barrier is: the metal film that plates the about 1mm of diameter in the centre of common K9 glass window.The thick more effect that is in the light of metal film is good more.If gold-plated film, when the thickness of gold was 100nm, the transmitance of zero level hot spot was less than 1%.Constantly change the position of each ligh trap through spatial light modulator, just can the figure that bead is arranged be transformed into smoothly the figure of Fig. 2 (d) in partly through a few minutes in Fig. 2 (b) part.Fig. 2 (c) part time space light modulator modulates is for this reason given the phase diagram of laser.
Regulating the 8th lens 18 is imaged on the Position-Sensitive Detector back focal plane of back object lens 15.Catoptron 17 passes through the anti-infrared laser of visible light.Laser intensity on the back focal plane of back object lens 15 distributes and has reacted the position of ligh trap and the position that bead departs from the ligh trap center.Back object lens 15, the eight lens 18, Position-Sensitive Detector 24 has constituted micro-displacement measuring system.The back focal plane interferometric method of this measurement micro-displacement has detailed introduction in list of references " Gittes; F.&Schmidt; C.F (1998) .Interference model for back-focal-plane displacement detection in optical tweezers.Optical letters 23:7-9 ", this paper repeats no more.
The optical maser wavelength that present embodiment adopts is 1064nm.A little less than the light absorption very of biological living sample to 1064nm, this has just been avoided the damage of the laser of strong convergence to biological sample.The particle size that the light tweezer can be caught is between tens nanometers to tens micron, and the size of a lot of tiny organism bodies such as cell, organelle is just also in this scope.Handle particulate because the light tweezer is the light with " invisible ", thereby the damage of having avoided Mechanical Contact that biosome is caused, and the environment around the biosome that is hunted down can not receive any interference, this has just guaranteed the normal vital movement of biosome.Therefore, this cover multiple light forceps system is particularly suitable for doing the biological living experiment.
In addition, for convenient forward and backward object lens are moved, guarantee the degree of accuracy and the degree of stability of system, the primary optical axis of present embodiment (being the residing optical axises of forward and backward object lens) is and is horizontally disposed with, and promptly primary optical axis is parallel to surface level.
It should be noted last that above embodiment is only unrestricted in order to technical scheme of the present invention to be described.Although the present invention is specified with reference to embodiment; Those of ordinary skill in the art is to be understood that; Technical scheme of the present invention is made amendment or is equal to replacement, do not break away from the spirit and the scope of technical scheme of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (6)
1. a three-dimensional light tweezers system comprises: laser output unit, spatial light modulator, laser convergence unit, sample cell and subjective measurement unit;
Said spatial light modulator is used for the single beam laser of said laser output unit output is divided into multiple laser;
Said laser convergence unit comprises convergence unit lens combination and preceding object lens, and said convergence unit lens combination is used for each Shu Jiguang of spatial light modulator output is expanded bundle, said before object lens be used for respectively restrainting laser convergence to said sample cell behind the bundle with expanding;
Said subjective measurement unit comprises visible light source, back object lens and subjective measurement unit CCD,
The optical axis conllinear of said visible light source, preceding object lens, back object lens and subjective measurement unit CCD forms primary optical axis; Said sample cell between preceding object lens and back object lens and with said before object lens all adjacent with back object lens; Object lens all can move along said primary optical axis with the back object lens before said, and said visible light source is positioned on the primary optical axis of preceding object lens one side.
2. three-dimensional light tweezers system according to claim 1; It is characterized in that; Said laser convergence unit also comprises convergence unit CCD and the semi-transparent semi-reflecting level crossing of convergence unit; The semi-transparent semi-reflecting level crossing of said convergence unit is used for the light path of assembling unit CCD is coupled into said primary optical axis between visible light source and preceding object lens.
3. three-dimensional light tweezers system according to claim 1; It is characterized in that; Said laser output unit is used to export the laser of infrared band, and said convergence unit also comprises the anti-infrared visible level crossing of convergence unit, is used for the light path of said convergence unit lens combination is coupled into said primary optical axis.
4. three-dimensional light tweezers system according to claim 1; It is characterized in that; Said laser output unit is used to export the laser of infrared band; Said three-dimensional light tweezers system also comprises small location measurement unit, and said small location measurement unit comprises the anti-infrared visible level crossing of the location measurement unit between back object lens and subjective measurement unit CCD, and the light path of said small location measurement unit is coupled into said primary optical axis through the anti-infrared visible level crossing of said location measurement unit.
5. three-dimensional light tweezers system according to claim 1 is characterized in that, said laser output unit is used for the laser that output wavelength is 1064nm.
6. three-dimensional light tweezers system according to claim 1 is characterized in that said primary optical axis is parallel to surface level.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102998293A (en) * | 2012-12-20 | 2013-03-27 | 武汉大学 | Multichannel quantitative detection device and detection method of two-photon fluorescence optical tweezers |
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| C06 | Publication | ||
| PB01 | Publication | ||
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