CN110082282A - The method and apparatus for realizing optical ultra-discrimination imaging based on optical tweezer - Google Patents
The method and apparatus for realizing optical ultra-discrimination imaging based on optical tweezer Download PDFInfo
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- CN110082282A CN110082282A CN201910314337.2A CN201910314337A CN110082282A CN 110082282 A CN110082282 A CN 110082282A CN 201910314337 A CN201910314337 A CN 201910314337A CN 110082282 A CN110082282 A CN 110082282A
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- 230000003287 optical effect Effects 0.000 title claims abstract description 34
- 238000003384 imaging method Methods 0.000 title claims abstract description 26
- 238000012576 optical tweezer Methods 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims description 8
- 238000005286 illumination Methods 0.000 claims abstract description 19
- 238000006073 displacement reaction Methods 0.000 claims abstract description 8
- 230000008878 coupling Effects 0.000 claims abstract description 7
- 238000010168 coupling process Methods 0.000 claims abstract description 7
- 238000005859 coupling reaction Methods 0.000 claims abstract description 7
- 230000000694 effects Effects 0.000 claims abstract description 5
- 230000005540 biological transmission Effects 0.000 claims abstract description 3
- 238000005259 measurement Methods 0.000 claims description 11
- 239000004005 microsphere Substances 0.000 claims description 2
- 239000013307 optical fiber Substances 0.000 claims description 2
- 239000000243 solution Substances 0.000 description 5
- 230000006378 damage Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 3
- 239000000835 fiber Substances 0.000 description 2
- 238000000399 optical microscopy Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 240000001307 Myosotis scorpioides Species 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241000294142 Vascellum Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000010287 polarization Effects 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000012846 protein folding Effects 0.000 description 1
- 230000006432 protein unfolding Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/32—Micromanipulators structurally combined with microscopes
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/361—Optical details, e.g. image relay to the camera or image sensor
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
- G02B27/58—Optics for apodization or superresolution; Optical synthetic aperture systems
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N2015/1481—Optical analysis of particles within droplets
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- Optics & Photonics (AREA)
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Abstract
Optical ultra-discrimination imaging method is realized based on optical tweezer the present invention relates to one kind and device, device include the displacement platform for placing sample, sample surfaces are marked with the solution with microballoon, laser, beam expander, Piezoelectric Driving reflecting mirror, lighting source, dichroscope, semi-transparent semi-reflecting lens and camera.The directional light of laser emitting expands laser by beam expander;Laser after expanding is reflected via Piezoelectric Driving reflecting mirror, the rotation center of Piezoelectric Driving reflecting mirror is overlapped via the laser of Piezoelectric Driving reflecting mirror reflection with incident light axis center and is coupled into same optical path by dichroscope by the illumination light that lighting source is emitted, and wherein the effect of dichroscope is transmission laser and indirect illumination light;Light after coupling enters object lens through semi-transparent semi-reflecting lens, and ligh trap is formed after the laser of object lens focuses for capturing microballoon.
Description
Technical field
The present invention relates to the method and apparatus for realizing optical ultra-discrimination imaging based on optical tweezer.
Technical background
Optical tweezer is formed by three-dimensional potential well using the laser beam of height convergence to stablize the micron that capture is located near focal point
Magnitude particle, spatial resolution, the mechanics resolution ratio of skin ox magnitude and non-contact, undamaged spy with Nano grade
Point is widely used in the fields such as cell biology, unimolecule biology, physics and colloid science, capture and behaviour by scholars
It controls the cell dredging vascellum blocking in intravital mouse ear capillary, capture polystyrene/silica dioxide microballoon and then studies
Protein unfolding/folding kinetics characteristic etc..However the above research is all based on capture and the manipulation micron level grain of optical tweezer
Son, optical tweezer, which is widely applied, should not be limited to thoses mentioned above, therefore realize that the technology of super-resolution needs to be probed into using optical tweezer.
Microsphere lens may be implemented super-resolution imaging, and can with a variety of optical microscopies, such as it is fluorescence, non-fluorescence aobvious
Micro mirror, wide field, spot scan microscope, is used in combination and keeps the hyperresolution of itself, for auxiliary optical microscopy imaging
And improve resolution ratio and provide a kind of simple method, it has broad application prospects.But how to place and move microballoon is
It realizes that a difficult point of this technology, general method are to be clamped and moved microballoon using mechanical arm, reuses object lens to see
Examine micro-structure.This to realize the system complex of super-resolution and expensive, and one can be realized the machinery of nanoscale mobile accuracy
Arm configuration is complicated and cost is very high, but also can generate injury to measurement sample.
Current commonly used optical tweezer measuring system and super-resolution measuring system all have the defects that many and deficiency:
(1) in optical optical tweezers system, the sample for needing to measure generally all is observed using transmitted light, is caused many opaque
Sample is not available optical tweezer and measures, and greatly reduces the application range of optical tweezer.And the measurement environment of optical tweezer is typically all
In the solution, for some samples that can not be fabricated to sample cell, optical tweezer also cannot achieve effective measurement.
(2) in super-resolution imaging system, the fixation and movement of microballoon are generally all realized using mechanical arm, leads to system
Structure is complicated, and with the raising of precision, the cost of system can also be increased substantially.In the process of fixed and mobile microballoon
In, mechanical arm can squeeze microballoon, and deformation occurs, generates error so as to cause measurement result.
Summary of the invention
In view of the above-mentioned problems, providing a kind of operation the purpose of the present invention is combining super-resolution imaging technology and optical tweezer technology
Flexibly simple, precision is higher and the method and apparatus of realization super-resolution imaging that will not damage to sample.Skill of the invention
Art scheme is as follows:
A kind of device for realizing optical ultra-discrimination imaging based on optical tweezer, including the displacement platform for placing sample, sample table
Face is marked with the solution with microballoon, laser, beam expander, Piezoelectric Driving reflecting mirror, lighting source, and dichroscope is semi-transparent semi-reflecting
Mirror and camera, wherein
The directional light of laser emitting expands laser by beam expander;
Laser after expanding is reflected via Piezoelectric Driving reflecting mirror, the rotation center and incident light axis of Piezoelectric Driving reflecting mirror
Center is overlapped
It is coupled via the laser of Piezoelectric Driving reflecting mirror reflection and by the illumination light of lighting source outgoing by dichroscope
Into same optical path, wherein the effect of dichroscope is transmission laser and indirect illumination light;
Light after coupling enters object lens through semi-transparent semi-reflecting lens, and ligh trap is formed after the laser of object lens focuses for capturing
Microballoon, illumination light are used for optical imagery;
Reflected light reflects after returning to object lens through semi-transparent semi-reflecting lens, and the light beam after reflection is reached camera and carried out by optical filter
Imaging, optical filter only allow illumination light to pass through to filter laser.
Preferably, laser is the fiber coupling solid-state laser of 1064nm wavelength continuous wave output;The object lens are
The object lens of high-NA.
Present invention simultaneously provides the optical ultra-discrimination imaging methods that above-mentioned device is realized:
1) it loads sample and captures microballoon using ligh trap, microballoon is placed on to the micro-structure surface for needing to measure, is then made
Image is obtained with micro-imaging optical path, fine tuning is realized by moving displacement platform or by the mobile ligh trap of Piezoelectric Driving reflecting mirror, makes
Obtain the microballoon that ligh trap captures sample surfaces;
2) the surface texture image of sample different location is obtained by mobile example and microballoon position;
3) image measured in different location is spliced by merging algorithm for images, obtains measurement knot to the end
Fruit.
The present invention proposes a kind of new measurement method and device, with prior art for the difficult point of measurement nano-micro structure
Compared to having the advantage that
(1) it focuses to form ligh trap capture microballoon using the collimation laser of height convergence, and passes through rotary piezoelectric ceramic reflecting
Mirror moves the microballoon captured, and accurately microballoon can be moved to above the micro-structure for needing to measure, and enormously simplifies previous
The experimental provision of microballoon is moved using mechanical arm, it may have higher precision, operation is more flexible, and is contactless survey
Amount, so will not be damaged to microballoon, to will not influence measurement result.
(2) it is 1064 nanometers of laser that wavelength has been used in the present apparatus, so will not damage and lead to sample to be tested
Cause measurement result inaccurate;Wavelength is that 1064 nanometers of laser is black light, so capture laser will not influence imaging system
Image quality, so as to preferably measure sample to be tested.
(3) imaging is amplified to sample to be tested using reflected light in the present apparatus, so compared in the past most of
Transparent sample can only be measured in optical optical tweezers system, the present apparatus can measure opaque sample, and application range is more extensive.
(4) the measurement environment of sample to be tested can be also possible in air in a liquid.For it is some can not be in Breakup of Liquid Ring
The sample measured under border can be measured directly in air.
(5) most of component in the present apparatus is all made of normal component, does not need to carry out additional processing, be easily installed,
It can also be replaced after parts damages, to extend system service life.
Detailed description of the invention
Fig. 1 is the overall structure figure of the device of the invention that optical ultra-discrimination imaging is realized based on optical tweezer.
Figure label explanation:
1: laser 2: beam expander 3: Piezoelectric Driving reflecting mirror 4:LED lamp
5: object lens 6: sample 7: displacement platform 8: optical filter
9: semi-transparent semi-reflecting lens 10:CCD camera 11: dichroscope
Specific embodiment:
Of the invention is described in detail with reference to the accompanying drawings and examples.
As shown in Figure 1, a kind of device that optical ultra-discrimination imaging is realized based on optical tweezer, it is characterized in that:
Laser 1 used in system is the fiber coupling solid-state laser 1 of 1064nm wavelength continuous wave output, laser
The light of 1 outgoing is directional light, power stability 2%~5%, the small 0.25mm of laser beam position stability.Optical fiber-coupled laser
It is that the laser head separated at one with laser pump (ing) excites, the temperature change of laser head is smaller, therefore the laser generated is usual
With extraordinary position stability.
The directional light that laser 1 is emitted expands laser perpendicular through the center of beam expander 2.Laser after expanding
It is reflected via Piezoelectric Driving reflecting mirror 3, and the rotation center of Piezoelectric Driving reflecting mirror 3 will be overlapped with incident light axis center.Swash
Light and the illumination light being emitted by LED light 4 pass through dichroscope 11 and are coupled into same optical path, and the wherein effect of dichroscope 11 is
Penetrate laser and indirect illumination light.Light after coupling enters object lens 5 through semi-transparent semi-reflecting lens 9, and laser forms ligh trap after focusing and is used for
Bead is captured, illumination light is used for optical imagery.Sample 6 is fixed on displacement platform 7, can be with mobile example by control bit moving stage 7
6.Reflected light reflects after returning to object lens 5 through semi-transparent semi-reflecting lens 9, the light beam after reflection by optical filter 8, reach CCD camera 10 into
Row imaging.Wherein the effect of optical filter 8 is to filter laser, only illumination light is allowed to pass through to protect CCD camera 10.
The method and apparatus for realizing super-resolution based on optical tweezer include following equipment and operation:
Step A: the collimation laser of height convergence and the object lens 5 of high-NA being emitted using laser 1 form one
The optical tweezer of high Optical Trap Stiffness, for capturing and mobile microballoon.
The optical tweezer of big Optical Trap Stiffness is formed by the object lens 5 of high-NA using the collimated laser beam of height convergence,
The laser beam is linear polarization Gaussian beam, the beam expander 2 being made of by one two lens, so having biggish light intensity
Gradient.
Step B: a set of visible light micro-imaging optical path suitable for optical optical tweezers system is built using CCD camera 10, using saturating
It penetrates light and carries out reflection micro-imaging.
Use LED light 4 as lighting source, optical tweezer optical path and illumination path have a part of some coincidences, in the present invention
A dichroscope 11 and semi-transparent semi-reflecting lens 9 have been used, have been individually positioned in front of the lighting source incidence object lens 5 and outgoing object
After mirror 5, for changing the direction of illumination light, and it is used to coupled illumination light and laser before by sample 6, is passing through sample
Separation incident light and emergent light after 6.Emergent light after separation only passes through illumination light, illumination light using an optical filter 8
For micro-imaging.
Step C: the solution of microballoon is had using the fixed sample to be tested 6 of displacement platform 7, and in sample surfaces injection.
If measuring environment is liquid environment, the solution of microballoon is just had in the drop of sample to be tested surface, then
It measures.If necessary to measure in air, need first to wait solution evaporation clean, then carry out subsequent measurement.
Step D: loading sample 6 and captures microballoon using optical tweezer, microballoon is placed on to the micro-structure surface for needing to measure, so
Image is obtained using micro-imaging optical path afterwards, passes through the surface of available 6 different location of sample of mobile example 6 and microballoon position
Structural images.
By moving displacement platform 7, the microballoon of sample surfaces is captured using optical tweezer, if it is the mobile microballoon of small distance,
It can be realized by the mobile ligh trap of Piezoelectric Driving reflecting mirror 3;If necessary to a wide range of mobile microballoon, then need to put by movement
Microballoon is placed on 6 surface of sample for needing to measure by the sample 6 set on displacement platform 7.Since microballoon realizes that super-resolution is not
In the focal point of object lens 5, so ligh trap position is not overlapped with imaging position, so temporary close laser is needed, then with aobvious
The micro-structure under microballoon is imaged in micro- imaging optical path, and the available size of CCD camera 10 is less than micro- knot of diffraction limit
The shape appearance figure of structure.
Step E: selecting the microballoon of different size and material to measure, finally obtain best imaging effect, to
The image arrived carries out post-processing.
The microballoon that diameter does not wait be can choose to measure, obtain the different image of resolution ratio, microsphere diameter gets over great achievement
Picture region is bigger, and the smaller imaging resolution of microballoon is higher.It is also an option that the microballoon of different materials, generally there are commonly titanium dioxides
The refractive index of silicon microballoon, different materials is different, so having a great impact to image quality.Measurement solution also can choose difference
Liquid have an impact since different solution refractive index are different to the size and imaging of trapping stiffness.It finally needs to spell by image
It connects algorithm to splice the image measured in different location, obtains measurement result to the end.
Claims (5)
1. a kind of device for realizing optical ultra-discrimination imaging based on optical tweezer, including the displacement platform for placing sample, sample surfaces
It is marked with the solution with microballoon, laser, beam expander, Piezoelectric Driving reflecting mirror, lighting source, dichroscope, semi-transparent semi-reflecting lens
And camera, wherein
The directional light of laser emitting expands laser by beam expander;
Laser after expanding is reflected via Piezoelectric Driving reflecting mirror, the rotation center of Piezoelectric Driving reflecting mirror and incident light axis center
It is overlapped
It is coupled into together via the laser of Piezoelectric Driving reflecting mirror reflection and by the illumination light that lighting source is emitted by dichroscope
One optical path, wherein the effect of dichroscope is transmission laser and indirect illumination light;
Light after coupling enters object lens through semi-transparent semi-reflecting lens, and it is micro- for capturing that ligh trap is formed after the laser of object lens focuses
Ball, illumination light are used for optical imagery;
Reflected light reflects after returning to object lens through semi-transparent semi-reflecting lens, and the light beam after reflection is reached camera and be imaged by optical filter,
Optical filter only allows illumination light to pass through to filter laser.
2. the apparatus according to claim 1, which is characterized in that laser is the optical fiber coupling of 1064nm wavelength continuous wave output
Close solid-state laser.
3. the apparatus according to claim 1, which is characterized in that the object lens are the object lens of high-NA.
4. as follows using the optical ultra-discrimination imaging method that device described in claim 1 is realized:
1) it loads sample and captures microballoon using ligh trap, microballoon is placed on to the micro-structure surface for needing to measure, then using aobvious
Micro- imaging optical path obtains image, fine tuning is realized by moving displacement platform or by the mobile ligh trap of Piezoelectric Driving reflecting mirror, so that light
Trap captures the microballoon of sample surfaces;
2) the surface texture image of sample different location is obtained by mobile example and microballoon position;
3) image measured in different location is spliced by merging algorithm for images, obtains measurement result to the end.
5. according to the method described in claim 4, being divided it is characterized in that, selection diameter not equal microballoon measures
The different image of resolution, microsphere diameter more big imaging region is bigger, and the smaller imaging resolution of microballoon is higher.
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Cited By (6)
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CN110955107A (en) * | 2019-11-18 | 2020-04-03 | 长春理工大学 | Ultra-high-speed time resolution camera shooting device and method based on reflection imaging technology |
CN111982812A (en) * | 2020-08-18 | 2020-11-24 | 天津大学 | Method for realizing optical super-resolution imaging by utilizing micron-scale liquid drops generated in real time |
CN113156634A (en) * | 2021-04-07 | 2021-07-23 | 华侨大学 | Multi-scene real-time application ultra-portable optical tweezers |
CN113484322A (en) * | 2021-07-13 | 2021-10-08 | 天津大学 | Optical tweezers super-resolution imaging method and system capable of feeding back axial optical trap position in real time |
CN113502207A (en) * | 2021-08-18 | 2021-10-15 | 长春长光辰英生物科学仪器有限公司 | Multifunctional cell sorting device based on laser system and operation method |
CN114077168A (en) * | 2022-01-06 | 2022-02-22 | 之江实验室 | Super-resolution laser direct writing and real-time imaging device and method based on optical tweezers microspheres |
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CN108469686A (en) * | 2018-06-20 | 2018-08-31 | 大连理工大学 | A kind of optical ultra-discrimination rate imaging system based on optical tweezer and microsphere lens |
CN108917651A (en) * | 2018-07-06 | 2018-11-30 | 中国科学院光电技术研究所 | Super-resolution three-dimensional shape measurement method based on optical tweezers medium microspheres |
CN109239937A (en) * | 2018-09-15 | 2019-01-18 | 天津大学 | A kind of optical tweezer automation control device |
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CN104749105A (en) * | 2015-04-21 | 2015-07-01 | 武汉大学 | Quantitative detection device and detection method based on near-infrared optical tweezers excited up-conversion luminescence |
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Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
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CN110955107A (en) * | 2019-11-18 | 2020-04-03 | 长春理工大学 | Ultra-high-speed time resolution camera shooting device and method based on reflection imaging technology |
CN111982812A (en) * | 2020-08-18 | 2020-11-24 | 天津大学 | Method for realizing optical super-resolution imaging by utilizing micron-scale liquid drops generated in real time |
CN113156634A (en) * | 2021-04-07 | 2021-07-23 | 华侨大学 | Multi-scene real-time application ultra-portable optical tweezers |
CN113484322A (en) * | 2021-07-13 | 2021-10-08 | 天津大学 | Optical tweezers super-resolution imaging method and system capable of feeding back axial optical trap position in real time |
CN113484322B (en) * | 2021-07-13 | 2023-01-10 | 天津大学 | Optical tweezers super-resolution imaging method and system capable of feeding back axial optical trap position in real time |
CN113502207A (en) * | 2021-08-18 | 2021-10-15 | 长春长光辰英生物科学仪器有限公司 | Multifunctional cell sorting device based on laser system and operation method |
CN113502207B (en) * | 2021-08-18 | 2022-11-15 | 长春长光辰英生物科学仪器有限公司 | Multifunctional cell sorting device based on laser system and operation method |
CN114077168A (en) * | 2022-01-06 | 2022-02-22 | 之江实验室 | Super-resolution laser direct writing and real-time imaging device and method based on optical tweezers microspheres |
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Application publication date: 20190802 |