阅读说明：本技术 一种基于透明介质微球的超分辨多色激光扫描光纤探针及其制作方法 (Super-resolution multi-color laser scanning optical fiber probe based on transparent medium microspheres and manufacturing method thereof ) 是由 王宏达 邵丽娜 石岩 蔡明军 初宏亮 曹自然 于 2019-09-26 设计创作，主要内容包括：本发明提供一种基于透明介质微球的超分辨多色激光扫描光纤探针以及制作方法。该探针包括多色空间激光-光纤主动耦合稳束系统和透明介质微球；该系统采用多色空间激光-光纤主动耦合稳束系统将多色空间激光汇聚成一束光,耦合进入第一光纤端口；然后采用基于透明介质微球的扫描光纤探针制作装置将透明介质微球放置并固定在第二光纤端口的芯径中心上,多色激光耦合进入第一光纤端口,通过光纤将多色激光传递到第二光纤端口处,经过透明介质微球后产生超分辨聚焦光斑。本发明的光纤探针,具有灵活、简单、稳定以及抑制杂散光等特点,解决了基于微球的超分辨率成像系统微球难操纵问题,为实现微球高速超分辨系统的二维或三维扫描成像提供了一种方法。(The invention provides a super-resolution multi-color laser scanning fiber probe based on transparent medium microspheres and a manufacturing method thereof. The probe comprises a multicolor space laser-optical fiber active coupling beam stabilizing system and transparent medium microspheres; the system adopts a multicolor space laser-optical fiber active coupling beam stabilizing system to converge multicolor space laser into a beam of light, and the beam of light is coupled into a first optical fiber port; and then, a scanning optical fiber probe manufacturing device based on the transparent medium microspheres is adopted to place and fix the transparent medium microspheres on the core diameter center of a second optical fiber port, the multi-color laser is coupled to enter the first optical fiber port, the multi-color laser is transmitted to the second optical fiber port through the optical fiber, and super-resolution focusing light spots are generated after passing through the transparent medium microspheres. The optical fiber probe has the characteristics of flexibility, simplicity, stability, stray light inhibition and the like, solves the problem that the microspheres of a microsphere-based super-resolution imaging system are difficult to operate, and provides a method for realizing two-dimensional or three-dimensional scanning imaging of a microsphere high-speed super-resolution system.)

1. A super-resolution multi-color laser scanning fiber probe based on transparent medium microspheres is characterized by comprising a multi-color space laser-fiber active coupling beam stabilizing system and transparent medium microspheres;

the multicolor space laser-optical fiber active coupling beam stabilizing system comprises a blue space laser, a green space laser, a red space laser, a first half-wave plate, a second half-wave plate, a third half-wave plate, a first electric double-shaft movable reflector, a second electric double-shaft movable reflector, a third electric double-shaft movable reflector, a fourth electric double-shaft movable reflector, a fifth electric double-shaft movable reflector, a sixth electric double-shaft movable reflector and a first polarization beam splitter, the device comprises a first polarization beam splitter, a first four-quadrant photoelectric detector, a second four-quadrant photoelectric detector, a third four-quadrant photoelectric detector, a first dichroic mirror, a second dichroic mirror, a third dichroic mirror, an acousto-optic modulator, an achromatic coupling lens, a high-precision three-dimensional displacement table, a first optical fiber port and a second optical fiber port;

the blue space laser, the green space laser and the red space laser emit parallel linear polarized laser beams which respectively pass through a first half-wave plate, a second half-wave plate and a third half-wave plate and then are respectively incident to a first electric double-shaft movable reflector, a second electric double-shaft movable reflector, a third electric double-shaft movable reflector, a fourth electric double-shaft movable reflector, a fifth electric double-shaft movable reflector and a sixth electric double-shaft movable reflector, then the light respectively reaches a first polarization beam splitter, a second polarization beam splitter and a third polarization beam splitter, the first polarization beam splitter, the second polarization beam splitter and the third polarization beam splitter respectively split a blue space laser, a green space laser and a red space laser into two parts, and one part of the light respectively enters a first four-quadrant photodetector, a second four-quadrant photodetector and a third four-quadrant photodetector; the other part of light respectively enters a first dichroic mirror, a second dichroic mirror and a third dichroic mirror, the light is combined through the first dichroic mirror, the second dichroic mirror and the third dichroic mirror, the combined laser is incident into an acousto-optic modulator and then enters an achromatic coupling lens, the achromatic coupling lens converges parallel light, the achromatic coupling lens is fixed on a high-precision three-dimensional displacement table, the combined laser is coupled and enters a first optical fiber port through adjusting the high-precision three-dimensional displacement table, and the first optical fiber port transmits the combined laser to a second optical fiber port through optical fibers;

the transparent medium microspheres are fixed on the core diameter of the second optical fiber port.

2. The transparent dielectric microsphere-based super-resolution multicolor laser scanning fiber-optic probe according to claim 1, wherein the blue space laser comprises a laser wavelength range of 400-492 nm; the green space laser comprises a laser wavelength range of 492-597 nm; the red space laser comprises a laser wavelength range of 597-1100 nm.

3. The super-resolution multicolor laser scanning fiber probe based on the transparent medium microspheres according to claim 1, characterized in that the optical fiber is a single mode fiber or a multimode fiber, the core diameter range of the single mode fiber is 4-10 micrometers, and the core diameter range of the multimode fiber is 10-100 micrometers.

4. The method for manufacturing the super-resolution multi-color laser scanning fiber probe based on the transparent dielectric microsphere according to claim 1, wherein the size of the transparent dielectric microsphere is 1-200 μm.

5. The method for manufacturing the super-resolution multi-color laser scanning fiber-optic probe based on the transparent dielectric microsphere according to claim 1, which comprises the following steps:

step one, building a multicolor space laser-optical fiber active coupling beam stabilizing system;

step two, forming the transparent medium microsphere solution into a transparent medium microsphere single-layer film by utilizing an automatic loading technology, then placing the optical fiber on a glass substrate, fixing a second optical fiber port on a digital microscope operating platform, fixing a first micro-tube holder by using a three-dimensional adjustable micro-operating machine, adsorbing the transparent medium microspheres on the glass substrate by using a first micro-tube on the first micro-tube holder, placing the transparent medium microspheres on the upper part of the center of the second optical fiber port, then a second micro-tube on a second micro-tube holder fixed on the right hand of the three-dimensional adjustable micro-manipulator machine adsorbs the ultraviolet light curing optical adhesive, the ultraviolet light curing optical adhesive is placed on a quartz interface around a fiber core of a second optical fiber port, and (3) under a digital microscope, irradiating by using an ultraviolet light source, and fixing the transparent medium microspheres on the core diameter of the second optical fiber port to obtain the super-resolution multi-color laser scanning optical fiber probe based on the transparent medium microspheres.

6. The method for manufacturing the super-resolution multi-color laser scanning fiber probe based on the transparent dielectric microsphere of claim 5, wherein the transparent dielectric microsphere solution is obtained by diluting a transparent dielectric microsphere material in an ethanol solution and performing ultrasonic treatment by using an ultrasonic instrument.

7. The method according to claim 6, wherein the transparent microsphere material comprises quartz, polystyrene or barium titanate.

8. The method for manufacturing a super-resolution multi-color laser scanning fiber probe based on transparent medium microspheres according to claim 5, wherein the first microtube on the first microtube holder is connected with a microinjector control box, and the microinjector control box generates positive or negative pressure on the first microtube.

9. The method for manufacturing the super-resolution multi-color laser scanning fiber probe based on the transparent dielectric microsphere of claim 5, wherein the negative pressure of the first microtube for adsorbing the transparent dielectric microsphere is 10-30 hPa.

10. The method for manufacturing the super-resolution multi-color laser scanning fiber-optic probe based on the transparent dielectric microspheres of claim 5, wherein the surface power density of the ultraviolet light source is 30mw/cm²～50mw/cm²The exposure time was two minutes.

Technical Field

The invention relates to the field of microscopic particle super-resolution microscopic imaging, in particular to a super-resolution multicolor laser scanning optical fiber probe based on transparent medium microspheres and a manufacturing method thereof.

Background

The resolution of a conventional optical microscope is generally not more than half the wavelength according to the rayleigh criterion because of the far-field diffraction limit. Therefore, for visible light imaging, the lateral resolution of a conventional optical microscope is typically only about 230 nm. In 2011 Wang et al reported microsphere lens imaging techniques to achieve 50nm resolution under white light. The simple and effective technology provides a new possibility for real-time super-resolution imaging.

The current microsphere-based super-resolution imaging system has the main principle that: the method comprises the steps of paving a microsphere on the surface of an observation sample, generating a focusing light spot smaller than a diffraction limit near the backlight surface of the microsphere and at several wavelengths due to a photon nanometer jetting effect, and receiving reflection imaging of the sample by using a traditional microscope. The regulation and the optimization of the nanometer focusing light spot can be realized by changing the laser wavelength, the size of the medium microsphere and the refractive index.

The microsphere-based super-resolution imaging system has certain defects, because microspheres are not easy to manipulate, observation positions are random, the field angle is small, and scanning imaging in two-dimensional or three-dimensional directions is difficult to realize. Meanwhile, the microspheres are directly irradiated by white light or laser, so that the problem of serious background light exists, and the imaging contrast is low.

Disclosure of Invention

The invention aims to solve the problems that the conventional microsphere is difficult to manipulate and inhibit background light, and provides a super-resolution multicolor laser scanning fiber-optic probe based on a transparent medium microsphere and a manufacturing method thereof.

The invention firstly provides a super-resolution multicolor laser scanning optical fiber probe based on transparent medium microspheres, which comprises a multicolor space laser-optical fiber active coupling beam stabilizing system and transparent medium microspheres;

the multicolor space laser-optical fiber active coupling beam stabilizing system comprises a blue space laser, a green space laser, a red space laser, a first half-wave plate, a second half-wave plate, a third half-wave plate, a first electric double-shaft movable reflector, a second electric double-shaft movable reflector, a third electric double-shaft movable reflector, a fourth electric double-shaft movable reflector, a fifth electric double-shaft movable reflector, a sixth electric double-shaft movable reflector and a first polarization beam splitter, the device comprises a first polarization beam splitter, a first four-quadrant photoelectric detector, a second four-quadrant photoelectric detector, a third four-quadrant photoelectric detector, a first dichroic mirror, a second dichroic mirror, a third dichroic mirror, an acousto-optic modulator, an achromatic coupling lens, a high-precision three-dimensional displacement table, a first optical fiber port and a second optical fiber port;

the blue space laser, the green space laser and the red space laser emit parallel linear polarized laser beams which respectively pass through a first half-wave plate, a second half-wave plate and a third half-wave plate and then are respectively incident to a first electric double-shaft movable reflector, a second electric double-shaft movable reflector, a third electric double-shaft movable reflector, a fourth electric double-shaft movable reflector, a fifth electric double-shaft movable reflector and a sixth electric double-shaft movable reflector, then the light respectively reaches a first polarization beam splitter, a second polarization beam splitter and a third polarization beam splitter, the first polarization beam splitter, the second polarization beam splitter and the third polarization beam splitter respectively split a blue space laser, a green space laser and a red space laser into two parts, and one part of the light respectively enters a first four-quadrant photodetector, a second four-quadrant photodetector and a third four-quadrant photodetector; the other part of light respectively enters a first dichroic mirror, a second dichroic mirror and a third dichroic mirror, the light is combined through the first dichroic mirror, the second dichroic mirror and the third dichroic mirror, the combined laser is incident into an acousto-optic modulator and then enters an achromatic coupling lens, the achromatic coupling lens converges parallel light, the achromatic coupling lens is fixed on a high-precision three-dimensional displacement table, the combined laser is coupled and enters a first optical fiber port through adjusting the high-precision three-dimensional displacement table, and the first optical fiber port transmits the combined laser to a second optical fiber port through optical fibers;

the transparent medium microspheres are fixed on the core diameter of the second optical fiber port.

Preferably, the blue space laser comprises a laser wavelength range of 400-492 nm; the green space laser comprises a laser wavelength range of 492-597 nm; the red space laser comprises a laser wavelength range of 597-1100 nm.

Preferably, the optical fiber is a single-mode optical fiber or a multi-mode optical fiber, the core diameter range of the single-mode optical fiber is 4-10 micrometers, and the core diameter range of the multi-mode optical fiber is 10-100 micrometers.

Preferably, the size of the transparent medium microsphere is 1-200 microns.

The invention also provides a method for manufacturing the super-resolution multicolor laser scanning optical fiber probe based on the transparent medium microspheres, which comprises the following steps:

step one, building a multicolor space laser-optical fiber active coupling beam stabilizing system;

step two, forming the transparent medium microsphere solution into a transparent medium microsphere single-layer film by utilizing an automatic loading technology, then placing the optical fiber on a glass substrate, fixing a second optical fiber port on a digital microscope operating platform, fixing a first micro-tube holder by using a three-dimensional adjustable micro-operating machine, adsorbing the transparent medium microspheres on the glass substrate by using a first micro-tube on the first micro-tube holder, placing the transparent medium microspheres on the upper part of the center of the second optical fiber port, then a second micro-tube on a second micro-tube holder fixed on the right hand of the three-dimensional adjustable micro-manipulator machine adsorbs the ultraviolet light curing optical adhesive, the ultraviolet light curing optical adhesive is placed on a quartz interface around a fiber core of a second optical fiber port, and (3) under a digital microscope, irradiating by using an ultraviolet light source, and fixing the transparent medium microspheres on the core diameter of the second optical fiber port to obtain the super-resolution multi-color laser scanning optical fiber probe based on the transparent medium microspheres.

Preferably, the transparent medium microsphere solution is obtained by diluting a transparent medium microsphere material in an ethanol solution and performing ultrasonic treatment by using an ultrasonic instrument.

Preferably, the transparent dielectric microsphere material comprises quartz, polystyrene or barium titanate.

Preferably, the first microtube in the first microtube holder is connected to a microinjector control box which generates a positive or negative pressure on the first microtube.

Preferably, the negative pressure intensity of the first microtube for adsorbing the transparent medium microspheres is 10-30 hPa.

Preferably, the surface power density of the ultraviolet light source is 30mw/cm²～50mw/cm²The exposure time was two minutes.

The invention has the advantages of

The invention provides a super-resolution multicolor laser scanning fiber probe based on transparent medium microspheres and a manufacturing method thereof, wherein the probe comprises a multicolor space laser-fiber active coupling beam stabilizing system and transparent medium microspheres, the multicolor space laser-fiber active coupling beam stabilizing system couples multicolor laser space laser into optical fibers, two high-precision electric double-shaft movable reflectors are adopted to form a high-precision electric double-shaft movable reflector set, a four-quadrant photoelectric detector forms a closed-loop control system, the laser spot position is automatically adjusted at high precision, and the active beam stability control and high-precision adjustment of the multicolor space laser-fiber coupling system are realized; the half-wave plate and the polarization beam splitter are adopted to form a proportional beam splitter, the distribution of laser energy with different proportions is realized by rotating the half-wave plate, and the laser energy is respectively used for being incident to a four-quadrant photoelectric detector to realize a laser spot position detection branch and entering an optical fiber coupling branch; the method comprises the steps that a dichroic mirror is utilized to combine multi-color lasers, the combined lasers are incident into an acousto-optic modulator and then enter an achromatic coupling lens, parallel light is converged by the achromatic coupling lens, the achromatic coupling lens is fixed on a high-precision three-dimensional displacement table, the combined lasers are coupled and enter a first optical fiber port through precision adjustment of the high-precision three-dimensional displacement table, and then the combined lasers enter a second optical fiber port through optical fibers.

Compared with the prior art, the invention can be applied to two-dimensional and three-dimensional scanning optical microscopes, reduces stray light and improves contrast; the super-resolution scanning optical fiber probe based on the transparent medium microspheres can form a focusing light spot with super-diffraction limit, and can be used as a light source for nano lithography or nano sensing. By adopting the super-resolution multicolor laser scanning optical fiber probe based on the transparent medium microspheres, one probe can meet the requirements on different wavelengths, and the super-resolution fluorescence imaging with specificity on the labeled biological samples in the field of life science can be met. The super-resolution multicolor laser scanning optical fiber probe based on the transparent medium microspheres and the manufacturing method can be used for researching the fields of biological super-resolution imaging, nano lithography, nano sensing and the like.

Drawings

FIG. 1 is a schematic structural diagram of a super-resolution multi-color laser scanning fiber probe based on transparent medium microspheres according to the present invention;

FIG. 2 is an enlarged view of a second optical fiber port of the present invention with immobilized transparent dielectric microspheres;

FIG. 3 is an image of a transparent dielectric microsphere monolayer film produced by the self-assembly technique under an optical microscope in accordance with the present invention;

FIG. 4 is a schematic view of a microscope micro-operation device of the super-resolution multi-color laser scanning fiber probe based on transparent medium microspheres in the manufacturing process of the invention;

in the figure, 1, blue space laser, 2, green space laser, 3, red space laser, 4, first half-wave plate, 5, second half-wave plate, 6, third half-wave plate, 7, first electric two-axis movable mirror, 8, second electric two-axis movable mirror, 9, third electric two-axis movable mirror, 10, fourth electric two-axis movable mirror, 11, fifth electric two-axis movable mirror, 12, sixth electric two-axis movable mirror, 13, first polarization beam splitter, 14, second polarization beam splitter, 15, third polarization beam splitter, 16, first four quadrant photodetector, 17, second four quadrant photodetector, 18, third four quadrant photodetector, 19, first dichroic mirror, 20, second dichroic mirror, 21, third dichroic mirror, 22, acousto-optic modulator, 23, achromatic coupling lens, 24. 25 parts of a high-precision three-dimensional displacement platform, 25 parts of a first optical fiber port, 26 parts of a second optical fiber port, 27 parts of a transparent dielectric microsphere single-layer film, 28 parts of a glass substrate, 29 parts of a digital microscope operating platform, 30 parts of a three-dimensional adjustable micro-operation mechanical left hand, 31 parts of a first micro-tube clamp, 32 parts of a transparent dielectric microsphere, 33 parts of a three-dimensional adjustable micro-operation mechanical right hand, 34 parts of a second micro-tube clamp, 35 parts of an ultraviolet light curing optical adhesive, 36 parts of a digital microscope.

Detailed Description

The invention firstly provides a super-resolution multicolor laser scanning optical fiber probe based on transparent medium microspheres, which comprises a multicolor space laser-optical fiber active coupling beam stabilizing system and transparent medium microspheres 32 as shown in a figure 1-2;

the multicolor space laser-optical fiber active coupling beam stabilizing system comprises a blue space laser 1, a green space laser 2, a red space laser 3, a first half-wave plate 4, a second half-wave plate 5, a third half-wave plate 6, a first electric double-shaft movable reflector 7, a second electric double-shaft movable reflector 8, a third electric double-shaft movable reflector 9, a fourth electric double-shaft movable reflector 10, a fifth electric double-shaft movable reflector 11, a sixth electric double-shaft movable reflector 12, a first polarization beam splitter 13, a second polarization beam splitter 14, a third polarization beam splitter 15, a first four-quadrant photoelectric detector 16, a second four-quadrant photoelectric detector 17, a third four-quadrant photoelectric detector 18, a first dichroic mirror 19, a second dichroic mirror 20, a third dichroic mirror 21, an acousto-optic modulator 22, an achromatic coupling lens 23, A high-precision three-dimensional displacement stage 24, a first fiber port 25 and a second fiber port 26;

the blue space laser 1, the green space laser 2 and the red space laser 3 emit parallel linear polarized laser beams, the parallel linear polarized laser beams respectively pass through a first half-wave plate 4, a second half-wave plate 5 and a third half-wave plate 6 and then respectively enter a first electric double-shaft movable reflector 7, a second electric double-shaft movable reflector 8, a third electric double-shaft movable reflector 9, a fourth electric double-shaft movable reflector 10, a fifth electric double-shaft movable reflector 11 and a sixth electric double-shaft movable reflector 12, and then respectively reach a first polarization beam splitter 13, a second polarization beam splitter 14 and a third polarization beam splitter 15, the first polarization beam splitter 13, the second polarization beam splitter 14 and the third polarization beam splitter 15 respectively split the blue space laser 1, the green space laser 2 and the red space laser 3 into two parts, and one part of the two parts respectively enter a first fourth quadrant photoelectric detector 16, a second quadrant photoelectric detector 16, a third photoelectric detector and a third photoelectric detector, A second fourth quadrant photodetector 17 and a third fourth quadrant photodetector 18; the other part of light respectively enters a first dichroic mirror 19, a second dichroic mirror 20 and a third dichroic mirror 21, the light is combined through the first dichroic mirror 19, the second dichroic mirror 20 and the third dichroic mirror 21, the combined laser is incident into an acousto-optic modulator 22 and then enters an achromatic coupling lens 23, the achromatic coupling lens 23 converges parallel light, the achromatic coupling lens 23 is fixed on a high-precision three-dimensional displacement table 24, the combined laser is coupled and enters a first optical fiber port 25 through adjusting the high-precision three-dimensional displacement table 24, and the first optical fiber port 25 transmits the combined laser into a second optical fiber port 26 through an optical fiber;

the transparent dielectric microspheres 32 are fixed on the core diameter of the second optical fiber port 26.

The invention also provides a method for manufacturing the super-resolution multicolor laser scanning optical fiber probe based on the transparent medium microspheres, which mainly comprises two steps as shown in figures 3-4: firstly, a multicolor space laser-optical fiber active coupling beam stabilizing system is built, and multicolor laser space laser is coupled into an optical fiber; secondly, fixing a single transparent medium microsphere on the optical fiber by adopting a microscope micro-operation system, which comprises the following steps

Step one, constructing a multicolor space laser-optical fiber active coupling beam stabilizing system: the blue space laser 1, the green space laser 2 and the red space laser 3 emit parallel linear polarized laser beams, the parallel linear polarized laser beams respectively pass through a first half-wave plate 4, a second half-wave plate 5 and a third half-wave plate 6 and then respectively enter a first electric double-shaft movable mirror 7, a second electric double-shaft movable mirror 8, a third electric double-shaft movable mirror 9, a fourth electric double-shaft movable mirror 10, a fifth electric double-shaft movable mirror 11 and a sixth electric double-shaft movable mirror 12, and then respectively reach a first polarization beam splitter 13, a second polarization beam splitter 14 and a third polarization beam splitter 15, the first polarization beam splitter 13, the second polarization beam splitter 14 and the third polarization beam splitter 15 respectively split the blue space laser 1, the green space laser 2 and the red space laser 3 into two parts, and one part of the light respectively enters a first four-quadrant photoelectric detector 16, A second fourth quadrant photodetector 17 and a third fourth quadrant photodetector 18; the other part of light respectively enters a first dichroic mirror 19, a second dichroic mirror 20 and a third dichroic mirror 21, the light is combined through the first dichroic mirror 19, the second dichroic mirror 20 and the third dichroic mirror 21, the combined laser is incident into an acousto-optic modulator 22 and then enters an achromatic coupling lens 23, the achromatic coupling lens 23 converges parallel light, the achromatic coupling lens 23 is fixed on a high-precision three-dimensional displacement table 24, the combined laser is coupled and enters a first optical fiber port 25 through precisely adjusting the high-precision three-dimensional displacement table 24, and the first optical fiber port 25 transmits the combined laser into a second optical fiber port 26 through optical fibers;

step two: dripping a trace amount of transparent medium microsphere solution into a container filled with a surfactant solution, standing for more than ten hours, wherein the transparent medium microspheres are automatically filled into a transparent medium microsphere single-layer film 27, then taking the cleaned and hydrophilically treated glass substrate, inclining the glass substrate by about 30 degrees, inserting the glass substrate below the liquid level, and slowly lifting upwards, wherein the transparent medium microsphere single-layer film 27 is transferred onto the glass substrate 28 as shown in FIG. 3; the transparent medium microspheres can be single transparent medium microspheres or transparent medium microsphere arrays; the size range of the transparent medium microspheres is 1-200 microns;

then the second optical fiber port 26 is fixed on the operation table 29 of the digital microscope, as shown in fig. 3, a three-dimensional adjustable micro-operation machine left hand 30 is used for fixing a first micro-tube holder 31, the first micro-tube holder 31 holds the first micro-tube, the first micro-tube is connected with a micro-injection instrument control box, the micro-injection instrument control box can generate positive or negative pressure to the first micro-tube, the first micro-tube uses negative pressure to adsorb one transparent medium microsphere 32 on the glass substrate 28, the micro-operation machine left hand 30 is operated to place the microsphere on the upper center of the second optical fiber port 26, then a second micro-tube holder 34 fixed on a three-dimensional adjustable micro-operation machine right hand 33 is used for clamping the second micro-tube 34, the second micro-tube adsorbs a small amount of ultraviolet light curing optical adhesive 35 of the micro-tube, and places the ultraviolet light curing optical adhesive 35 on the quartz interface around the fiber, the pressure of the first micro-tube fixed on the left hand 30 of the micro-operation machine is regulated and controlled to become zero, the transparent medium microspheres 32 adsorbed by the first micro-tube are placed on the fiber core of the second optical fiber port 26, and the ultraviolet light source is used for irradiating and curing under the digital microscope 36, so that the super-resolution multi-color laser scanning optical fiber probe based on the transparent medium microspheres is obtained.

In the first step, the blue space laser 1 includes a laser wavelength range of 400-492 nm; the green space laser 2 comprises a laser wavelength range of 492-597 nm; the red space laser 3 comprises a laser wavelength range of 597-1100 nm;

in the first step, the first half-wave plate 4, the second half-wave plate 5 and the third half-wave plate 6 are used for changing the linear polarization angles of the emergent lasers of the blue space laser 1, the green space laser 2 and the red space laser 3, so that the polarization angles of the three beams of lasers are consistent, and the coupling efficiency is highest;

in the first step, the first electric double-shaft movable reflecting mirror 7, the second electric double-shaft movable reflecting mirror 8, the third electric double-shaft movable reflecting mirror 9, the fourth electric double-shaft movable reflecting mirror 10, the fourth electric double-shaft movable reflecting mirror 11 and the sixth electric double-shaft movable reflecting mirror 12 are respectively used as reflecting mirror groups, and each reflecting mirror group can realize high-precision angle adjustment and position adjustment of laser. The angle adjustment comprises pitch angle adjustment and torsion angle adjustment, and the position adjustment comprises horizontal direction adjustment and vertical direction adjustment;

in the first step, the variation of the four-quadrant laser intensity of the first four-quadrant photodetector 16, the second four-quadrant photodetector 17, and the third four-quadrant photodetector 18 reflects the position deviation of the blue space laser 1, the green space laser 2, and the red space laser 3, respectively. The four quadrant laser intensity on the four quadrant photodetector is recorded as A, B, C, D, respectively. The up-down offset amount may be represented as (a + B) - (C + D), and the left-right offset amount may be represented as (a + D) - (B + C). A first four-quadrant photodetector 16, a second four-quadrant photodetector 17 and a third four-quadrant photodetector 18 respectively form closed-loop control with the reflector groups of the first electric double-shaft movable reflector 7, the second electric double-shaft movable reflector 8, the third electric double-shaft movable reflector 9, the fourth electric double-shaft movable reflector 10, the fourth electric double-shaft movable reflector 11 and the sixth electric double-shaft movable reflector 12;

in the first step, the first dichroic mirror 19, the second dichroic mirror 20, and the third dichroic mirror 21 are long-wave pass filters, respectively, and the cut-off wavelength of the filters is related to that the first dichroic mirror 19< the second dichroic mirror 20< the third dichroic mirror 21;

in the first step, the numerical aperture of the achromatic coupling lens 23 is required to be matched with the numerical aperture of the first optical fiber port 25;

in the first step, the optical fiber can be a single-mode optical fiber and a multi-mode optical fiber, the core diameter of the single-mode optical fiber ranges from 4 micrometers to 10 micrometers, and the core diameter of the multi-mode optical fiber ranges from 10 micrometers to 100 micrometers.

In the second step, the transparent medium microsphere solution is obtained by diluting the transparent medium microspheres in an ethanol solution according to a volume ratio of 1:1, and performing ultrasonic treatment for 10 minutes by using an ultrasonic instrument.

In the second step, the transparent dielectric microspheres preferably comprise quartz, polystyrene or barium titanate.

In the second step, the glass substrate 28 is moved into a mixed solution of sulfuric acid and hydrogen peroxide according to the volume ratio of 3:1, soaked for more than half an hour, and then cleaned by ultrapure water to remove residual substances.

In the second step, the negative pressure of the first microtube on the first microtube holder 31 for adsorbing the transparent medium microspheres 32 is 10-30 hPa.

In the second step, the surface power density of the ultraviolet light source is 30mw/cm²～50mw/cm²The exposure time was two minutes.