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 PDF

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Publication number
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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laser
optical
microballoon
light
reflecting mirror
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胡春光
马彦华
张雅静
林祖增
胡小唐
胡晓东
李宏斌
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Tianjin University
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Tianjin University
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N15/14Optical investigation techniques, e.g. flow cytometry
    • G01N15/1468Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/32Micromanipulators structurally combined with microscopes
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/36Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/361Optical details, e.g. image relay to the camera or image sensor
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/58Optics for apodization or superresolution; Optical synthetic aperture systems
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N15/14Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1481Optical analysis of particles within droplets

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  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Optics & Photonics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
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  • Microscoopes, Condenser (AREA)
  • Length Measuring Devices By Optical Means (AREA)

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

The method and apparatus for realizing optical ultra-discrimination imaging based on optical tweezer
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.
CN201910314337.2A 2019-04-18 2019-04-18 The method and apparatus for realizing optical ultra-discrimination imaging based on optical tweezer Pending CN110082282A (en)

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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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Publication number Priority date Publication date Assignee Title
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
CN104777602A (en) * 2015-04-23 2015-07-15 东北大学 Device for classifying and collecting atmosphere PM2.5 particles through hollow optical fiber taper and optical tweezers
CN106485204A (en) * 2016-09-19 2017-03-08 天津大学 A kind of microballoon automatic capture method in optical optical tweezers system
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Cited By (8)

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Publication number Priority date Publication date Assignee Title
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