A method and apparatus for achieving micro-imaging under ultra-high-speed structured illumination

Abstract

This invention discloses a method for achieving ultra-high-speed structured illumination micro-imaging, comprising: 1) shaping an illumination beam emitted from a light source to form an illumination beam with uniform intensity distribution; 2) splitting the illumination beam into a central interference optical path and multiple interference sub-paths, each interference sub-path exhibiting illumination interference fringes in different directions on the sample surface; 3) performing optical path compensation on each interference sub-path to ensure that all sub-paths have the same optical path length, and that the light from each interference sub-path is split into p-polarized light and s-polarized light; 4) combining the beam from the central interference optical path with the p-polarized and s-polarized light from each interference sub-path and illuminating the sample to form a structured illumination pattern; 5) collecting the fluorescence excited by the sample for microscopic imaging. This invention also discloses a device for achieving ultra-high-speed structured illumination micro-imaging. This invention can significantly improve the modulation speed of the illumination light, thereby achieving ultra-high-speed structured illumination imaging.

Description

Technical Field

[0001] This invention belongs to the field of optical engineering, and specifically relates to a method and apparatus for realizing ultra-high-speed structured illumination micro-imaging. Background Technology

[0002] Optical microscopy is a method for probing the microscopic world. Due to its non-contact and non-destructive nature, it is widely used in biomedical research. However, the resolution of traditional optical microscopy methods cannot exceed the diffraction limit, and therefore cannot be used to observe microstructures below 200 nm.

[0003] Structured illumination microscopy is a super-resolution imaging technique that modulates the illumination light to obtain multiple images of a sample under different illuminations, and finally reconstructs a super-resolution image. Its theoretical resolution limit can reach half of the diffraction limit. While structured illumination microscopy can improve resolution, it sacrifices imaging speed. The reconstruction of each super-resolution image requires several original images captured under different illumination conditions. Therefore, the final imaging speed depends not only on the camera's shooting and reading speed but also on the modulation speed of the illumination light.

[0004] In the field of structured light microscopy, traditional illumination light modulation methods include grating mechanical movement, spatial light modulator modulation, and galvanometer scanning. Among them, grating mechanical movement has the slowest modulation speed. Spatial light modulator modulation can be divided into two categories: liquid crystal spatial light modulator (SLM) and digital micromirror device (DMD). The former is slower, while the latter has lower diffraction efficiency. Galvanometer scanning has the fastest speed, but it still cannot meet the imaging speed of kHz.

[0005] Patent application CN105487214A discloses a rapid three-dimensional super-resolution microscopy method, comprising: converting a laser beam into linearly polarized light after collimation; phase-modulating the linearly polarized light and converting it into circularly polarized light for projection onto the sample; and collecting signal light emitted from each scanning point of the sample; performing three-dimensional scanning of the sample; wherein the phase modulation includes primary phase modulation and secondary phase modulation; the primary phase modulation uses a spatial light modulator to phase-modulate the s-component of the linearly polarized light, and the secondary phase modulation uses a spatial light modulator to phase-modulate the p-component of the linearly polarized light; finally, a three-dimensional super-resolution image is obtained based on the effective signal light intensity. However, the use of a spatial light modulator for illumination light modulation results in a relatively slow imaging speed.

[0006] Patent application CN206848565U discloses an optical focusing enhancement system based on a digital micromirror device (DMM). A first optical fiber and a collimating lens are arranged after a laser. The laser beam is transmitted through the first optical fiber and then incident on the DMM through the collimating lens. A light block is placed at the side exit end of the DMM, and a beam-shrinking module is placed at the front exit end. A dichroic mirror is positioned in front of the DMM. After being reflected by the dichroic mirror, the beam passes through a scanning module and enters the microscope objective for focusing. The experimental sample is located on the focal plane of the microscope objective. The fluorescence excited within the experimental sample passes through the microscope objective and the scanning module, then through the dichroic mirror and is received by a light intensity detection module for light intensity detection. Using a DMM for illumination light modulation suffers from low diffraction efficiency.

[0007] It is evident that existing solutions all have their limitations. To achieve ultra-high-speed structured illumination micro-imaging, a completely new design and methodology are needed to meet the requirements of high-speed imaging. Summary of the Invention

[0008] The purpose of this invention is to provide a method for achieving ultra-high-speed structured light illumination micro-imaging. This method can significantly improve the modulation speed of the illumination light, thereby achieving ultra-high-speed (KHz level) structured light illumination imaging.

[0009] A method for achieving micro-imaging under ultra-high-speed structured illumination includes:

[0010] 1) Shape the illumination beam emitted by the light source to form an illumination beam with uniform intensity distribution;

[0011] 2) Split the illumination beam into a central interference path and multiple sub-interference paths, each sub-interference path representing illumination interference fringes in different directions on the sample surface;

[0012] 3) Perform optical path compensation on each interferometric sub-path to make the optical path of all sub-paths the same, and the light of each interferometric sub-path is divided into p-polarized light and s-polarized light;

[0013] 4) The p and s polarized light of each interferometer optical path are modulated by an electro-optic modulator to maintain the phase difference, and then the polarization direction is changed by a half-wave plate to make their polarization directions consistent.

[0014] 5) The beam from the central interference optical path is combined with the p-polarized and s-polarized beams from each interference sub-path and then illuminates the sample to form a structured light illumination pattern.

[0015] 6) Collect the fluorescence excited by the sample for microscopic imaging.

[0016] In this invention, an image acquisition module based on a high-speed rotating mirror is used for imaging, which can capture multiple images in a single exposure, enabling high-speed capture of experimental images and matching the speed of illumination modulation.

[0017] Preferably, the illumination beam is reflected and transmitted after passing through a polarization beam splitter. The reflected light enters the central interference optical path, and the transmitted optical path is then divided into multiple interference sub-paths in sequence.

[0018] In specific embodiments, there may be three interferometer optical paths, or it may be expanded to multiple paths as needed to achieve more special structured light illumination patterns, such as special crystal-type illumination.

[0019] Preferably, the selection of each interferometer sub-optical path is selectively controlled. Each time an interferometer sub-optical path is selected, an image is recorded. After all sub-optical paths are selected in sequence, a complete set of structured light illumination images is obtained.

[0020] Preferably, each interferometer optical path is simultaneously selected, and the beams from each interferometer optical path interfere together to generate a two-dimensional structured light illumination pattern.

[0021] In this invention, each optical path can be selected simultaneously, and the energy of each sub-optical path can be distributed in any proportion, so that the beams of each sub-optical path interfere together to produce a two-dimensional structured light illumination pattern, instead of being limited to a one-dimensional grating-type illumination pattern.

[0022] This invention also provides a device for realizing ultra-high-speed structured illumination micro-imaging, comprising a light source emitting an illumination beam, an image acquisition module for acquiring fluorescence emitted by a sample, and further comprising: arranged in the optical path of the illumination beam.

[0023] The first polarization beam splitter splits the illumination beam, the reflected light enters the central interference optical path, and the transmitted light is sequentially divided into multiple interference sub-optical paths;

[0024] The gating module, corresponding to each of the multiple interferometer optical paths, is used to control the gating state of each interferometer optical path;

[0025] The second polarization beam splitter located on each interferometer optical path splits the light from each interferometer optical path into p-polarized light and s-polarized light;

[0026] The deflection beam combining module is used to combine the beam from the central interference optical path with the p-polarized and s-polarized beams from each interference sub-path.

[0027] After beam combining, the light is applied to the sample to form a structured light illumination pattern and excite fluorescence, which is then imaged onto the image acquisition module.

[0028] In this invention, the illumination laser generated by the light source is split into two beams by a first polarization beam splitter. One beam enters the central interference path, while the other beam is selected by multiple gating modules, causing multiple interferometric sub-paths to sequentially open and close. In each interferometric sub-path, the beam passes sequentially through a high-speed phase adjustment device and an optical path compensation system. The optical path compensation system compensates for the optical path difference between the central interference path and the interferometric sub-paths, thus maintaining the coherence among all four interference paths. The high-speed phase adjustment device converts the beam into s-polarized and p-polarized light with equal energy and a specific phase difference. The polarization beam splitter separates these two polarized beams, and, in conjunction with a mirror and a half-wave plate, changes the spatial angle of the beam. The half-wave plate rotates the polarization directions of the two interference beams, aligning their polarization directions and maximizing the contrast of the interference result. After beam combining, all the light passes through a beam combining imaging system, illuminating the sample surface and forming a structured light illumination pattern at the focal plane of the objective lens. The excited fluorescence is reflected by a dichroic mirror and ultimately captured by a high-speed image acquisition module. At the same time, the 10:90 beam splitter also reflects 10% of the energy. The reflected light enters the spectrum observation path, passes through the lens, and forms a spectrum image of the interference surface on the camera, which is used to monitor the interference beam.

[0029] Preferably, the light source and the first polarization beam splitter are sequentially provided with: an acousto-optic modulator for high-speed on / off control of the beam; a beam shaping module for generating collimated light with uniform spatial intensity distribution; and a beam shrinking module for changing the beam diameter to conform to the working aperture of the electro-optic modulator.

[0030] Preferably, the gating module includes a polarization electro-optic modulator and a polarization beam splitter, and the central interference optical path has, in sequence: a first phase electro-optic modulator for phase modulation of the beam; a first polarization modulator for changing the polarization direction of the beam; and a first beam expander for beam expansion.

[0031] In this invention, the selection state of the sub-optical path is controlled at high speed by an electro-optic modulator. The electro-optic modulator is used to perform high-speed phase modulation on the interference beam, control its phase difference, and generate different illumination patterns, thus breaking through the speed limit of ordinary phase modulation methods. Linearly polarized illumination is used, and the polarization direction of the beam is adjusted at high speed by an electro-optic modulator. In conjunction with a polarization beam splitter, the on / off state of different sub-optical paths is controlled at high speed.

[0032] The polarization electro-optic modulator can change the polarization direction of light, thereby controlling its transmission or reflection at the polarization beam splitter. The reflected light will enter the first interferometer optical path, and the transmitted light will enter the next set of gating modules, and continue to be divided into the second interferometer optical path and the third interferometer optical path.

[0033] Preferably, each interferometer optical path has, in sequence: a first half-wave plate, used to change the polarization direction of the beam so that the s-component and p-component of the beam are equal; a second phase electro-optic modulator, used to perform phase modulation on the beam so that the s-component and p-component of the beam generate a relative phase difference; a second beam expander module, used to expand the beam; an optical path compensation module, used to compensate for the optical path difference between each interferometer optical path; and a third polarization beam splitter, used to split the beam into p-polarized light and s-polarized light.

[0034] By combining the spatial angle of the interference beams, a half-wave plate is used to change the polarization direction of the interference beams in order to obtain the maximum interference contrast, and all interference beams will be corrected for optical path difference through the optical path compensation module.

[0035] Preferably, the p-polarized light and s-polarized light enter the deflection and beam combining module after passing through the second half-wave plate. The deflection and beam combining module has a set of mirrors corresponding to the beam, p-polarized light and s-polarized light in the central interference optical path. The deflection and beam combining module deflects all interference beams and combines them in space.

[0036] In this invention, all optical paths are ultimately converged onto the same illumination path, and their interference regions overlap. A fixed set of mirrors is used to control the spatial angle of the interference beam.

[0037] In a further preferred embodiment, the combined beam illuminates the sample through the objective lens, and the fluorescence emitted by the sample is imaged onto the image acquisition module. The image acquisition module includes a rotating mirror and a camera. After the rotating mirror, there are multiple imaging optical paths. The fluorescence circulates through the multiple imaging optical paths in sequence and is finally imaged onto different areas of the camera.

[0038] In this invention, the gating module, high-speed phase modulation device, and high-speed camera are linked for control. Each time a sub-optical path is selected, the high-speed phase modulation device for that path performs a series of related phase modulations. Each time the phase changes, an image is recorded on the high-speed camera. Then, another sub-optical path is selected, and the phase control and camera capture operations are repeated. Once all sub-optical paths are sequentially selected, a complete set of structured light illumination images is captured by the camera. The above steps are repeated to obtain multiple sets of structured light illumination images.

[0039] The high-speed image module uses a high-speed rotating mirror for scanning, dividing the camera target surface into several imaging areas, allowing the camera to capture multiple fluorescence images in a single exposure.

[0040] In this invention, a spatial filter can be set on the spectral surface after the illumination beam is combined to filter out the light of the required frequency order and improve the contrast of the final illumination pattern.

[0041] Compared with the prior art, the advantages of the present invention are as follows:

[0042] 1) Polarization modulation is performed using an electro-optic modulator to control the on / off state of each sub-optical path, with a gating speed of up to several hundred kHz.

[0043] 2) Phase modulation is achieved by using an electro-optic modulator, enabling ultra-high-speed phase modulation of the interference beam. The modulation speed can reach hundreds of kHz, which is far higher than existing technical solutions. The imaging speed of the entire system will no longer be limited by illumination modulation.

[0044] 3) Using a fixed reflector to control the angle of the interference beam provides greater stability;

[0045] 4) By using a half-wave plate to change the polarization direction of the interference beam, the contrast of the interference image is improved.

[0046] 5) All illumination beams are combined and share the same illumination imaging optical path;

[0047] 6) The high-speed image acquisition module uses a high-speed rotating mirror for scanning, recording multiple images in a single exposure, which can further increase the acquisition speed by several times on the basis of the camera's maximum imaging speed. Attached Figure Description

[0048] Figure 1 This is an optical path diagram of the ultra-high-speed structure illumination micro-imaging device in an embodiment of the present invention;

[0049] Figure 2 This is a schematic diagram of the deflection beam combining module in an embodiment of the present invention;

[0050] Figure 3 This is a schematic diagram of the high-speed image acquisition module of the present invention. Detailed Implementation

[0051] Numerous specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and therefore the invention is not limited to the specific embodiments disclosed below.

[0052] The embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout.

[0053] use Figure 1 The device shown illustrates a method for achieving high-speed structured illumination micro-imaging as follows:

[0054] Laser 1 emits an illumination beam, which is coupled into single-mode fiber 4 after passing through acousto-optic modulator 2 and first lens 3. Acousto-optic modulator 2 can achieve high-speed on / off control of the beam. The laser exits from the end of single-mode fiber 4, and after passing through second lens 5, it becomes Gaussian collimated light. The collimated light is then transformed into uniform flat-top collimated light after passing through beam shaping module 6, and then beam-shrinking module 7, so that the beam diameter matches the entrance aperture of each electro-optic modulator.

[0055] Before entering the first polarization beam splitter 9, the polarization direction of the beam is rotated by the first half-wave plate 8, causing the light intensity to be reflected and transmitted at a certain ratio after passing through the first polarization beam splitter 9. The reflected light enters the central interference optical path. The light entering the central interference optical path passes sequentially through the second half-wave plate 10, the first phase electro-optic modulator 11, and the first polarization electro-optic modulator 12. The first phase electro-optic modulator 11 modulates the phase of the beam before it enters the first polarization modulator 12, changing the polarization direction of the beam. Subsequently, the beam is expanded by the first beam expander module 13. The light transmitted after passing through the first polarization beam splitter 9 enters the second polarization electro-optic modulator 14, where it is selectively converted into p-polarized light (or s-polarized light). The s-polarized light is reflected at the second polarization beam splitter 15 and enters the first interferometer optical path, passing sequentially through the third half-wave plate 16, the second phase electro-optic modulator 17, the second beam expander module 18, and the first optical path compensation module 19. The third half-wave plate 16 changes the polarization direction of the beam, making the s-component and p-component of the beam equal. After passing through the second phase electro-optic modulator, the S-component and P-component generate a relative phase difference, which is then amplified by the second beam expander module 18. The P-polarized light modulated by the second polarization electro-optic modulator 14 enters the third polarization electro-optic modulator 20, where it is selectively converted back to P-polarized light (or S-polarized light). Similarly, the S-polarized light enters the second interferometer optical path, passing sequentially through the fourth half-wave plate 22, the third phase electro-optic modulator 23, the third beam expander module 24, and the second optical path compensation module 25, thus ensuring that the S-component and P-component intensities are equal and have a constant phase difference. The P-polarized light modulated by the third polarization electro-optic modulator 20, after being reflected by the first reflector 26, enters the third interferometer optical path, passing sequentially through the fifth half-wave plate 27, the fourth phase electro-optic modulator 28, the fourth beam expander module 29, and the third optical path compensation module 30, ensuring that the S-component and P-component intensities are equal and have a constant phase difference.

[0056] The first optical path compensation module 19, the second optical path compensation module 25, and the third optical path compensation module 30 will compensate for the optical path difference between each interference optical path so that the optical path difference of all interference beams is within the laser coherence length.

[0057] The beam from the central interference optical path is deflected by the second mirror 31, the third mirror 32 and the fourth mirror 33, and then incident parallel to the deflection beam combining module 56.

[0058] The light in the first interferometer optical path is split into p-polarized light and s-polarized light by the fourth polarization beam splitter 34. The p-polarized light passes through the sixth half-wave plate 35, the fifth mirror 36 and the sixth mirror 37 and enters the deflection beam combining module 56. The s-polarized light passes through the seventh half-wave plate 38, the seventh mirror 39, the eighth mirror 40 and the ninth mirror 41 and enters the deflection beam combining module 56.

[0059] Similarly, the light in the second interferometer optical path is split into p-polarized light and s-polarized light by the fifth polarization beam splitter 42. The p-polarized light passes through the eighth half-wave plate 43, the tenth mirror 44 and the eleventh mirror 45 and enters the deflection beam combining module 56; the s-polarized light passes through the ninth half-wave plate 46, the twelfth mirror 47 and the thirteenth mirror 48 and enters the deflection beam combining module 56.

[0060] Similarly, the light in the third interferometer optical path is split into p-polarized light and s-polarized light by the sixth polarization beam splitter 49. The p-polarized light passes through the tenth half-wave plate 50, the fourteenth mirror 51 and the fifteenth mirror 52 and enters the deflection beam combining module 56; the s-polarized light passes through the eleventh half-wave plate 53, the sixteenth mirror 54 and the seventeenth mirror 55 and enters the deflection beam combining module 56.

[0061] The deflection and beam combining module 56 deflects all interference beams at an appropriate angle and combines them spatially, finally projecting them onto the eighteenth reflecting mirror 57. After reflection, the beams pass through a 4f system composed of the third lens 58 and the fourth lens 62, where the Fourier surface has multiple interference orders. The 10:90 beam splitter 59 reflects 10% of the light, and the Fourier surface is imaged onto the pupil plane monitoring camera 61 by the fifth lens 63 for monitoring the pupil plane. The remaining 90% of the energy passes through the field lens 63, the fluorescence module 64, and the objective lens 65, illuminating the sample and forming a structured light illumination pattern. The three-dimensional translation sample stage can control the sample to perform three-dimensional translation, achieving lateral movement and axial scanning. The fluorescence emitted by the sample is reflected by the fluorescence module 64 and imaged onto the high-speed image acquisition module 68 by the sixth lens 67.

[0062] Figure 2 This is a schematic diagram of the deflection and beam combining module 56 of the present invention. The seven inclined reflective surfaces at specific angles can reflect the seven incident beams of light (one central interference beam and three pairs of sub-interference beams) upward at a certain angle, and the beams are deflected again by the upper reflector to achieve beam combining.

[0063] Figure 3This is a schematic diagram of the high-speed image acquisition module 68 of the present invention, including: a high-speed rotating mirror 69, a seventh lens 70, an eighth lens 71, a nineteenth reflecting mirror 72, a twentieth reflecting mirror 73, a ninth lens 74, a tenth lens 75, an eleventh lens 76, a twelfth lens 77, a twenty-first reflecting mirror 78, a twenty-second reflecting mirror 79, and a high-speed camera 80. The sample image plane is conjugate to the reflecting surface of the high-speed rotating mirror. Behind the high-speed rotating mirror are three imaging optical paths: a first imaging optical path (seventh lens 70, eighth lens 71, nineteenth reflecting mirror 72, twentieth reflecting mirror 73), a second imaging optical path (ninth lens 74, tenth lens 75), and a third imaging optical path (eleventh lens 76, twelfth lens 77, twenty-first reflecting mirror 78, twenty-second reflecting mirror 79). When the high-speed rotating mirror is working, fluorescence will sequentially circulate through these three imaging optical paths, ultimately being imaged onto different areas of the high-speed camera 80. A complete scan is completed within the time of a single exposure of 80 seconds using a high-speed camera, and three experimental images are captured simultaneously, which can further increase the acquisition speed by three times based on the camera's original maximum shooting speed.

[0064] In another embodiment, a method for realizing ultra-high-speed structured illumination micro-imaging is also provided, comprising:

[0065] 1) Shape the illumination beam emitted by the light source to form an illumination beam with uniform intensity distribution;

[0066] 2) The illumination beam is split into a central interference optical path and multiple interference sub-optical paths, each of which has illumination interference fringes in different directions on the sample surface;

[0067] 3) Perform optical path compensation on each interferometric sub-path to make the optical path of all sub-paths the same, and the light of each interferometric sub-path is divided into p-polarized light and s-polarized light;

[0068] 4) The beam from the central interference optical path is combined with the p-polarized and s-polarized beams from each interference sub-path and then illuminates the sample to form a structured light illumination pattern.

[0069] 5) Collect the fluorescence excited by the sample for microscopic imaging.

[0070] In this invention, the method can be implemented based on the above-described device embodiments, or it can be implemented using optical paths with other structures. Specifically, an image acquisition module based on a high-speed rotating mirror is used for imaging, enabling the formation of multiple images in a single exposure, high-speed acquisition of experimental images, and matching the speed of illumination modulation.

[0071] The illumination beam is reflected and transmitted after passing through a polarization beam splitter. The reflected light enters the central interference optical path, and the transmitted optical path is then divided into multiple interference sub-paths in sequence.

[0072] In other preferred embodiments, there may be three interferometer optical paths, or it may be expanded to multiple paths as needed to achieve more special structured light illumination patterns, such as special crystal-type illumination.

[0073] In this embodiment, the selection of each interferometric sub-path is selectively controlled. Each time an interferometric sub-path is selected, an image is recorded. After all sub-paths are selected in sequence, a complete set of structured light illumination images is obtained. Alternatively, all interferometric sub-paths can be selected simultaneously, and the beams from each interferometric sub-path interfere together to generate a two-dimensional structured light illumination pattern.

[0074] In this invention, each optical path can be selected simultaneously, and the energy of each sub-optical path can be distributed in any proportion, so that the beams of each sub-optical path interfere together to produce a two-dimensional structured light illumination pattern, instead of being limited to a one-dimensional grating-type illumination pattern.

[0075] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for achieving ultra-high-speed structured illumination micro-imaging, characterized in that, include: 1) Shape the illumination beam emitted by the light source to form an illumination beam with uniform intensity distribution; 2) Split the illumination beam into a central interference path and multiple sub-interference paths, each sub-interference path representing illumination interference fringes in different directions on the sample surface; The illumination beam is reflected and transmitted after passing through a polarization beam splitter. The reflected light enters the central interference optical path, and the transmitted optical path is then divided into multiple interference sub-paths in sequence. 3) Perform optical path compensation on each interferometric sub-path to make the optical path of all sub-paths the same, and the light of each interferometric sub-path is divided into p-polarized light and s-polarized light; 4) The p and s polarized light of each interferometer optical path are modulated by an electro-optic modulator to maintain the phase difference, and then the polarization direction is changed by a half-wave plate to make their polarization directions consistent. 5) The beam from the central interference optical path is combined with the p-polarized and s-polarized beams from each interference sub-path and then illuminates the sample to form a structured light illumination pattern. 6) Collect the fluorescence excited by the sample for microscopic imaging; The on / off state of each interferometer sub-path is selectively controlled. Each time an interferometer sub-path is selected, an image is recorded. After all sub-paths are selected in sequence, a complete set of structured light illumination images is obtained.

2. The method for achieving ultra-high-speed structured illumination micro-imaging according to claim 1, characterized in that, When all interferometer optical paths are simultaneously selected, the beams from each interferometer optical path interfere with each other, producing a two-dimensional structured light illumination pattern.

3. A device for realizing ultra-high-speed structured illumination micro-imaging, comprising a light source emitting an illumination beam and an image acquisition module for acquiring fluorescence emitted by a sample; characterized in that, It also includes those arranged in the optical path of the illumination beam: The first polarization beam splitter splits the illumination beam, the reflected light enters the central interference optical path, and the transmitted light is sequentially divided into multiple interference sub-optical paths; The light entering the central interference optical path passes sequentially through the second half-wave plate, the first phase electro-optic modulator, and the first polarization electro-optic modulator. The first phase electro-optic modulator modulates the phase of the beam, and the beam enters the first polarization modulator, changing the polarization direction of the beam. Subsequently, the first beam expansion module expands the beam. The light transmitted through the first polarization beam splitter enters the second polarization electro-optic modulator, where it is selectively converted into p-polarized or s-polarized light. The s-polarized light is reflected at the second polarization beam splitter and enters the first interferometer optical path. The p-polarized light modulated by the second polarization electro-optic modulator enters the third polarization electro-optic modulator, where it is again selectively converted into p-polarized or s-polarized light. The s-polarized light is reflected at the third polarization beam splitter and enters the second interferometer optical path, while the p-polarized light modulated by the third polarization electro-optic modulator enters the third interferometer optical path. The gating module, corresponding to each of the multiple interferometer optical paths, is used to control the gating state of each interferometer optical path; The deflection beam combining module is used to combine the beam from the central interference optical path with the p-polarized and s-polarized beams from each interference sub-path. After beam combining, the light is applied to the sample to form a structured light illumination pattern and excite fluorescence, which is then imaged onto the image acquisition module.

4. The device for realizing ultra-high-speed structured illumination micro-imaging according to claim 3, characterized in that, Between the light source and the first polarizing beam splitter, the following are sequentially arranged: Acousto-optic modulator, used for high-speed on / off control of light beams; A beam shaping module is used to generate collimated light with a uniform spatial intensity distribution; And a beam-shrinking module, used to change the beam diameter to match the operating aperture of the electro-optic modulator.

5. The device for realizing ultra-high-speed structured illumination micro-imaging according to claim 3, characterized in that, The gating module includes a polarization electro-optic modulator and a polarization beam splitter, with the following sequentially arranged on each interferometer optical path: The first half-wave plate is used to change the polarization direction of the beam so that the s-component and p-component of the beam are equal. The second phase electro-optic modulator is used to phase modulate the beam, and the s component and p component of the beam generate a relative phase difference. The second beam expander module is used for beam expansion. The optical path compensation module is used to compensate for the optical path difference between each interfering optical path; And a third polarization beam splitter, used to split the beam into p-polarized light and s-polarized light.

6. The device for realizing ultra-high-speed structured illumination micro-imaging according to claim 5, characterized in that, The p-polarized light and s-polarized light enter the deflection and beam combining module after passing through the second half-wave plate. The deflection and beam combining module has a set of mirrors corresponding to the beam, p-polarized light and s-polarized light in the central interference optical path. The deflection and beam combining module deflects all the interference beams and combines them in space.

7. The device for realizing ultra-high-speed structured illumination micro-imaging according to claim 3, characterized in that, The combined beam illuminates the sample through the objective lens, and the fluorescence emitted by the sample is imaged onto the image acquisition module. The image acquisition module includes a rotating mirror and a camera. After the rotating mirror, there are multiple imaging optical paths. The fluorescence passes through multiple imaging optical paths in sequence and is finally imaged onto different areas of the camera.

Images