Cell tissue morphology microscopic imaging method and imaging device thereof

Abstract

The invention relates to a cell tissue morphology microscopic imaging method and an imaging device thereof, solves the contradiction between the field of view and the resolution of a commercial microscope and the problem that only 2D information can be obtained, and belongs to the field of optical microscopic imaging. The method comprises the following steps: irradiating a to-be-detected 3D cell tissue sample in parallel by adopting a Fourier laminated illumination light beam, collecting diffracted light generated by the to-be-detected 3D cell tissue sample, dividing the diffracted light into two paths of optical signals according to a set proportion, obtaining low-resolution three-dimensional image information according to one path of optical signal, and obtaining two-dimensional image information according to the other path of optical signal; and performing three-dimensional reconstruction according to the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain a cell tissue morphology image of the to-be-detected 3D cell tissue sample, performing optical path fusion on a four-dimensional optical field and a Fourier lamination technology, and proposing three-dimensional reconstruction to realize large-view-field and high-resolution cell morphology imaging.

Description

Technical Field

[0001] The invention relates to a cell tissue morphology microscopic imaging method and an imaging device thereof, and belongs to the field of optical microscopic imaging. Background Art

[0002] 3D microscopy technology can provide 3D information at the micro- and nanoscale, allowing researchers to gain a deeper understanding of biological tissue and cell structure. This technology is widely used in many scientific fields, including biology, medicine, physics, and materials science.

[0003] At present, microscopic three-dimensional imaging technology has made significant progress. Existing three-dimensional microscopic imaging technologies include confocal microscopy, multiphoton microscopy, light sheet microscopy, three-dimensional microscopic imaging based on structured light, and three-dimensional microscopic imaging based on light field. High-resolution three-dimensional microscopic imaging can only be achieved by layer-by-layer scanning, which is inefficient and has a shallow depth of field; and direct three-dimensional imaging is difficult to improve resolution due to structural limitations.

[0004] In biomedical research, it is often necessary to culture certain cells in vitro in 3D. During the culture process, their status needs to be observed to study dynamic processes. Currently, commercial microscopes are mainly used for observation. However, commercial microscopes have low resolution when the field of view is large, and small field of view when the resolution is high. At the same time, they can only obtain 2D image information, which restricts the observation and research application of 3D cell culture process. Summary of the invention

[0005] In view of the contradiction between the field of view and resolution of commercial microscopes and the problem that only 2D information can be obtained, the present invention provides a cell tissue morphology microscopic imaging method and an imaging device thereof.

[0006] The cell tissue morphology microscopic imaging method of the present invention comprises:

[0007] A Fourier stacked illumination beam is used to parallely illuminate the 3D cell tissue sample to be tested, and the diffracted light generated by the 3D cell tissue sample to be tested is collected, and the diffracted light is divided into two optical signals according to a set ratio, and low-resolution three-dimensional image information is obtained according to one optical signal, and two-dimensional image information is obtained according to the other optical signal;

[0008] Three-dimensional reconstruction is performed based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain a cell tissue morphology image of the 3D cell tissue sample to be tested.

[0009] Preferably, the method of performing three-dimensional reconstruction based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain the cell tissue morphology image of the 3D cell tissue sample to be tested comprises:

[0010] Two models are constructed. The first model is a mathematical model that simulates the Fourier stacking process under a virtual aperture. The first model takes a three-dimensional point cloud as input and outputs a spectrum and phase map under different apertures. The second model is a mathematical model that simulates an optical sensor. The input is a three-dimensional point cloud and the output is a three-dimensional image.

[0011] The obtained low-resolution three-dimensional image information is used to construct three-dimensional point clouds with different focal lengths, and used as the initial input of the two models; the three-dimensional point cloud is used as the variable to be optimized, and regularization terms and constraints that conform to prior knowledge are added. The outputs of the two models are compared with the obtained low-resolution three-dimensional image information and two-dimensional image information, and the input three-dimensional point cloud is optimized. According to the optimized three-dimensional point cloud, the cell tissue morphology image of the 3D cell tissue sample to be tested is obtained.

[0012] Preferably, the method of using the Fourier stack illumination beam is:

[0013] Light sources at different positions are illuminated in a redundant and overlapping manner, and the diffracted light with high-frequency and low-frequency information is collected separately.

[0014] The present invention also provides an imaging device for the above-mentioned cell tissue morphology microscopic imaging method, comprising:

[0015] A light source module, used for parallelly irradiating a 3D cell tissue sample to be tested using a Fourier stacked illumination beam;

[0016] An acquisition module is used to collect diffracted light generated by the 3D cell tissue sample to be tested, and divide the diffracted light into two optical signals according to a set ratio, and obtain low-resolution three-dimensional image information according to one optical signal, and obtain two-dimensional image information according to the other optical signal;

[0017] The three-dimensional reconstruction module is used to perform three-dimensional reconstruction based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain a cell tissue morphology image of the 3D cell tissue sample to be tested.

[0018] Preferably, the light source module comprises a two-dimensional LED array and an eyepiece lens group;

[0019] The two-dimensional LED array adopts Fourier stack illumination, and the LED light of the two-dimensional LED array passes through the eyepiece lens group and then illuminates the 3D cell tissue sample to be tested in parallel.

[0020] Preferably, the light source control system lights up LEDs at different positions in the two-dimensional LED array in a time-sharing manner in an overlapping and redundant manner, so that the collection module collects the diffracted light with high-frequency and low-frequency information separately.

[0021] Preferably, the eyepiece lens group is a single lens group, a multi-lens group or a compound lens group.

[0022] Preferably, the acquisition module includes a microscope objective lens, a beam splitting prism, a two-dimensional image acquisition module and an optical sensor;

[0023] The microscope objective collects the diffracted light generated by the 3D cell tissue sample to be tested, and the diffracted light is incident on the beam splitter prism, which splits it into two light signals according to a set ratio, one of which enters the optical sensor and the other enters the two-dimensional image acquisition module;

[0024] The optical sensor obtains low-resolution three-dimensional image information according to one optical signal, and the two-dimensional image acquisition module obtains two-dimensional image information according to another optical signal.

[0025] Preferably, the optical sensor comprises a tube lens group No. 1, a microlens array and a camera No. 1;

[0026] One optical signal passes through tube lens group No. 1 and microlens array in sequence and is collected by camera No. 1. The image collected by camera No. 1 is low-resolution three-dimensional image information.

[0027] Preferably, the two-dimensional image acquisition module includes a No. 2 tube lens group and a No. 2 camera;

[0028] The other optical signal is collected by camera No. 2 through tube lens group No. 2, and the image collected by camera No. 2 is two-dimensional image information.

[0029] The beneficial effects of the present invention are as follows: the present invention uses Fourier stack illumination to improve the effective numerical aperture value in the resolution, and has a large field of view and high-resolution cell morphology imaging; uses light field microscopy imaging technology to obtain spatial position and angle information, realizes quantitative three-dimensional imaging, and provides an algorithm for three-dimensional reconstruction. The reconstruction method of the present invention is highly efficient, has low requirements for acquisition, and saves computing resources. The present invention does not require a precise and complex motion system and motion control system, and has the characteristics of low cost and simple structure. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is a schematic diagram of the principle of the cell tissue morphology microscopic imaging device;

[0031] Figure 2 It is a schematic diagram comparing the principles of traditional lighting and Fourier stacked lighting of the present invention;

[0032] Figure 3 Schematic diagram of the imaging principle of a traditional microscope system;

[0033] Figure 4 It is a schematic diagram of the three-dimensional imaging principle of the present invention. DETAILED DESCRIPTION

[0034] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0036] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but they are not intended to limit the present invention.

[0037] Specific implementation method 1: A cell tissue morphology microscopic imaging method of this implementation method includes:

[0038] A Fourier stacked illumination beam is used to parallely illuminate the 3D cell tissue sample to be tested, and the diffracted light generated by the 3D cell tissue sample to be tested is collected, and the diffracted light is divided into two optical signals according to a set ratio, and low-resolution three-dimensional image information is obtained according to one optical signal, and two-dimensional image information is obtained according to the other optical signal;

[0039] Three-dimensional reconstruction is performed based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain a cell tissue morphology image of the 3D cell tissue sample to be tested.

[0040] Traditional microscopes use a light beam with a certain aperture angle to illuminate the sample. The microscope objective collects the high-frequency and low-frequency information of the sample at the same time. The two are aliased and the low-frequency intensity is much greater than the high-frequency intensity, resulting in the inability to distinguish the high-frequency detail information of the sample. The Fourier stack illumination provided in this embodiment illuminates the sample at different positions at different times, collects the high-frequency and low-frequency information separately, and uses the provided high-frequency and low-frequency low-resolution images for three-dimensional reconstruction.

[0041] Traditional microscope systems can only obtain object position and intensity information on the camera plane and have a small imaging depth, which cannot meet the needs of application scenarios with 3D imaging requirements; or they can only achieve 3D reconstruction by stitching multiple layers through layer scanning, but this method requires precise and expensive focusing and motion systems. The light field imaging method reduces the 7-dimensional parameters (x, y, z, θ, φ, λ, t) in the traditional light field to 4 dimensions (s, w, u, v) by adding a group of microlens arrays at the focal plane position of the tube lens group. x, y, z represent the three-dimensional spatial position coordinates, θ represents the elevation angle of the light in the spatial coordinate system, φ represents the projection angle of the light in the spatial coordinate system, λ represents the wavelength, and t represents time; s represents the horizontal coordinate of the plane defining the position in the dual-plane space, w represents the vertical coordinate of the plane defining the position in the dual-plane space, u represents the horizontal coordinate of the plane defining the direction in the dual-plane space, and v represents the vertical coordinate of the plane defining the direction in the dual-plane space; (s, t, u, v) includes the sample information at different depths in the object space. The space formed by the plane where the microlens array is located and the detection plane can not only obtain the imaging surface position intensity information, but also the angle information, so as to reconstruct the images at different positions in the space using the refocusing algorithm based on the Fourier slice theorem to generate a three-dimensional image with quantitative depth.

[0042] This embodiment integrates the Fourier stack illumination technology with improved resolution and the 4D light field technology that can obtain quantitative depth information, thereby achieving quantitative 3D imaging function with large field of view and high resolution without the need for complex motion mechanisms.

[0043] The main function of Fourier stack-based microscopy is to improve the resolution of two-dimensional images. The three-dimensional reconstruction method of this embodiment combines the phase and amplitude obtained by the Fourier stack imaging method and the low-resolution three-dimensional image information collected by the light field sensor. After completing the acquisition of the original data of the sensor, the data is reconstructed and fused using the corresponding algorithm. In order to make full use of the fluorescence and light field image data of different depths of field to reconstruct a more accurate visualization effect of three-dimensional cell communication signals, in a preferred embodiment, in this embodiment, the method of performing three-dimensional reconstruction based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain the cell tissue morphology image of the 3D cell tissue sample to be tested includes:

[0044] Step 1, construct two models, the first model is a mathematical model for simulating the Fourier stacking process under a virtual aperture, the first model takes a three-dimensional point cloud as input, and takes the spectrum and phase map under different apertures as output, and the second model is a mathematical model for simulating an optical sensor, the input is a three-dimensional point cloud, and the output is a three-dimensional image; the microlens array 7 in the light field sensor is larger than the camera pixel, and a certain distance is left between the microlenses to avoid light aliasing, resulting in a low resolution of the three-dimensional imaging, so the three-dimensional point cloud generated in this step has a certain gap with the real three-dimensional structure;

[0045] Step 2: Use the obtained low-resolution three-dimensional image information to construct three-dimensional point clouds with different focal lengths, and use them as the initial input of the two models; use the three-dimensional point cloud as the variable to be optimized, and add regularization terms and constraints that conform to prior knowledge to ensure that the calculated three-dimensional point cloud has visual characteristics such as smoothness, continuity and spatial correlation. Use the outputs of the two models to compare with the acquired low-resolution three-dimensional image information and two-dimensional image information, optimize the input three-dimensional point cloud, and obtain the cell tissue morphology image of the 3D cell tissue sample to be tested based on the optimized three-dimensional point cloud. Due to the highly nonlinear and non-convex characteristics of the imaging model, it is difficult to directly use the optimization algorithm for rapid global optimization. Therefore, this embodiment uses the constructed three-dimensional point cloud with different focal lengths as the initial three-dimensional point cloud for optimization to avoid nonlinear optimization from falling into a local extreme value that is too far away from the real three-dimensional point cloud. The cell tissue morphology image of the 3D cell tissue sample to be tested is obtained based on the optimized three-dimensional point cloud.

[0046] The method of this embodiment uses Fourier stack illumination to improve the effective numerical aperture value in the resolution, and has a large field of view and high-resolution cell morphology imaging; uses light field microscopy imaging technology to obtain spatial position and angle information, realize quantitative three-dimensional imaging, and provide an algorithm for three-dimensional reconstruction. The reconstruction method of the present invention is highly efficient, has low requirements for acquisition, and saves computing resources. The present invention does not require a precise and complex motion system and motion control system, and has the characteristics of low cost and simple structure.

[0047] This embodiment also provides an imaging device based on the above-mentioned cell tissue morphology microscopic imaging method, comprising:

[0048] A light source module, used for parallelly irradiating a 3D cell tissue sample to be tested using a Fourier stacked illumination beam;

[0049] An acquisition module is used to collect diffracted light generated by the 3D cell tissue sample to be tested, and divide the diffracted light into two optical signals according to a set ratio, and obtain low-resolution three-dimensional image information according to one optical signal, and obtain two-dimensional image information according to the other optical signal;

[0050] The three-dimensional reconstruction module is used to perform three-dimensional reconstruction based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain a cell tissue morphology image of the 3D cell tissue sample to be tested.

[0051] In a preferred embodiment, the light source module in this embodiment includes a two-dimensional LED array 1 and an eyepiece lens group 2;

[0052] The 3D cell tissue sample to be tested is placed in the culture dish 3, and the two-dimensional LED array 1 adopts Fourier stack illumination. Preferably, the LEDs at different positions in the two-dimensional LED array 1 are illuminated in a time-sharing manner in an overlapping and redundant manner through the light source control system, so that the acquisition module collects the diffracted light with high-frequency and low-frequency information separately. The LED light of the two-dimensional LED array 1 passes through the wide-angle eyepiece lens group 2 and then irradiates the 3D cell tissue sample to be tested in the culture dish 3 in parallel, providing a uniform plane wave.

[0053] Traditional microscopes use a beam of light with a certain aperture angle to illuminate the sample. The microscope objective collects high-frequency and low-frequency information of the sample at the same time. The two are aliased and the low-frequency intensity is much greater than the high-frequency intensity, resulting in the inability to distinguish the high-frequency detail information of the sample. The Fourier stack illumination provided in this embodiment lights up the LEDs at different positions to illuminate the sample at different times, and collects the high-frequency and low-frequency information separately. The three-dimensional reconstruction method provided later uses the provided high-frequency and low-frequency low-resolution images for reconstruction. Figure 2 Schematic diagram of traditional lighting and Fourier stacked lighting of the present invention (the left side is traditional lighting, and the right side is lighting only one of the lights).

[0054] The acquisition module of this embodiment includes a microscope objective lens 4, a beam splitting prism 5, a two-dimensional image acquisition module and an optical sensor;

[0055] The microscope objective 4 collects the diffracted light generated by the 3D cell tissue sample to be tested, and the diffracted light is incident on the beam splitter prism 5, which splits the light into two optical signals according to a set ratio, which can be divided into two optical signals in a 1:1 manner or in other ratios. One of the two optical signals enters the optical sensor, and the other enters the two-dimensional image acquisition module;

[0056] The optical sensor obtains low-resolution three-dimensional image information according to one optical signal, and the two-dimensional image acquisition module obtains two-dimensional image information according to another optical signal.

[0057] In a preferred embodiment, the optical sensor of this embodiment includes tube lens group No. 1 6 - 1 , microlens array 7 and camera No. 1 8 - 1 ; microlens array 7 is located at the focal plane position of tube lens group No. 1 6 , and camera No. 1 8 - 1 is located at the focal plane position of microlens array 7 .

[0058] One of the two optical signals passes through tube lens group 6-1 and microlens array 7 in sequence, and is then collected by camera 8-1. The image collected by camera 8-1 is low-resolution three-dimensional image information. Camera 8-1 will collect a series of low-resolution original images. Different low-resolution images correspond to different sub-spectrum areas of the sample. The images are transmitted to the host computer and the resolution is improved using a three-dimensional reconstruction method.

[0059] In a preferred embodiment, the two-dimensional image acquisition module of this embodiment includes a No. 2 tube lens group 6 - 2 and a No. 2 camera 8 - 2 , and the No. 2 camera 8 - 2 is located at the focal plane position of the No. 2 tube lens group 6 .

[0060] The other optical signal is collected by the No. 2 camera 8 - 2 through the No. 2 tube lens group 6 - 2 , and the image collected by the No. 2 camera 8 - 2 is a two-dimensional image.

[0061] In conventional microscopy, the system resolution Where λ is the wavelength of the illumination source, NA obj The resolution of the microscope system is determined only by the objective lens used. where NA ill is the numerical aperture of the illumination system. From the formula, it can be seen that the system resolution is greatly improved due to the introduction of the illumination aperture, thereby achieving high-resolution imaging while maintaining the original microscope objective field of view, solving the contradiction between the field of view and resolution of the traditional microscope system.

[0062] like Figure 3 As shown, traditional microscope systems can only obtain object position intensity information on the camera plane and have a small imaging depth, which cannot meet the needs of application scenarios with 3D imaging requirements; or can only achieve 3D reconstruction by multi-layer stitching through layer scanning, but this method requires precise and expensive focusing and motion systems. The light field imaging method reduces the 7-dimensional parameters (x, y, z, θ, φ, λ, t) in the traditional light field to 4 dimensions (s, w, u, v) by adding a group of microlens arrays at the focal plane position of the tube lens group. x, y, z represent the three-dimensional spatial position coordinates, θ represents the elevation angle of the light in the spatial coordinate system, φ represents the projection angle of the light in the spatial coordinate system, λ represents the wavelength, and t represents time; s represents the horizontal coordinate of the plane defining the position in the dual-plane space, w represents the vertical coordinate of the plane defining the position in the dual-plane space, u represents the horizontal coordinate of the plane defining the direction in the dual-plane space, and v represents the vertical coordinate of the plane defining the direction in the dual-plane space; (s, t, u, v) includes sample information at different depths in the object space. The space formed by the plane where the microlens array 7 is located and the plane where the camera is located can not only obtain the intensity information of the imaging surface position, but also the angle information, so as to reconstruct images at different positions in the space using the refocusing algorithm based on the Fourier slice theorem to generate a three-dimensional image with quantitative depth, such as Figure 4 shown.

[0063] Based on the above-mentioned cell tissue morphology microscopic imaging device, the Fourier stack illumination technology with improved resolution and the 4-dimensional light field technology that can obtain quantitative depth information are optically fused, thereby realizing quantitative three-dimensional imaging function with a large field of view and high resolution without the need for complex motion mechanisms.

[0064] The main function of microscopic imaging based on Fourier stacking is to improve the resolution of two-dimensional images. On this basis, the three-dimensional reconstruction method of this embodiment combines the phase and amplitude obtained by the Fourier stacking imaging method and the three-dimensional image information with lower resolution collected by the light field sensor. After completing the collection of the original data of the sensor, the data is reconstructed and fused by the corresponding algorithm. In order to make full use of the fluorescence and light field image data of different depths of field to reconstruct a more accurate visualization effect of three-dimensional cell communication signals, in a preferred embodiment, the three-dimensional reconstruction module of this embodiment is used to construct two models. The first model is a mathematical model that simulates the Fourier stacking process under a virtual aperture. The first model takes a three-dimensional point cloud as input and outputs a spectrum and phase map under different apertures. The second model is a mathematical model that simulates an optical sensor, with a three-dimensional point cloud as input and a three-dimensional image as output; the microlens array 7 in the light field sensor is larger than the camera pixel, and a certain distance is left between the microlenses to avoid light aliasing, resulting in a low resolution of three-dimensional imaging. Therefore, the three-dimensional point cloud generated in this step has a certain gap with the real three-dimensional structure;

[0065] The three-dimensional reconstruction module of this embodiment is also used to construct three-dimensional point clouds of different focal lengths using the obtained low-resolution three-dimensional image information, and use them as the initial input of the two models; the three-dimensional point cloud is used as the variable to be optimized, and regular terms and constraints that conform to prior knowledge are added to ensure that the calculated three-dimensional point cloud has visual characteristics such as smoothness, continuity and spatial correlation. The output of the two models is compared with the obtained low-resolution three-dimensional image information and two-dimensional image information, and the input three-dimensional point cloud is optimized. According to the optimized three-dimensional point cloud, the cell tissue morphology image of the 3D cell tissue sample to be tested is obtained. Due to the highly nonlinear and non-convex characteristics of the imaging model, it is difficult to directly use the optimization algorithm for rapid global optimization. Therefore, this embodiment uses the constructed three-dimensional point cloud of different focal lengths as the initial three-dimensional point cloud for optimization to avoid nonlinear optimization from falling into a local extreme value that is too far away from the real three-dimensional point cloud. According to the optimized three-dimensional point cloud, the cell tissue morphology image of the 3D cell tissue sample to be tested is obtained.

[0066] This implementation has a large field of view and high resolution - Fourier stacked illumination improves the effective numerical aperture value in the resolution; it realizes quantitative three-dimensional imaging - light field microscopy technology, obtains spatial position and angle information, and uses algorithms to perform three-dimensional reconstruction; it has the characteristics of low cost and simple structure - it does not require precise and complex motion systems and motion control systems; the three-dimensional reconstruction module of this implementation is highly efficient, has low acquisition requirements, and saves computing resources.

[0067] Although the present invention is described herein with reference to specific embodiments, it should be understood that these embodiments are merely examples of the principles and applications of the present invention. It should therefore be understood that many modifications may be made to the exemplary embodiments and that other arrangements may be devised without departing from the spirit and scope of the present invention as defined by the appended claims. It should be understood that the various dependent claims and features described herein may be combined in a manner different from that described in the original claims. It should also be understood that features described in conjunction with individual embodiments may be used in other described embodiments.

Claims

1. A cell tissue morphology microscopic imaging method, characterized in that: The method comprises: Using a Fourier stacked illumination beam to illuminate the 3D cell tissue sample to be tested in parallel, collecting diffracted light generated by the 3D cell tissue sample to be tested, dividing the diffracted light into two light signals according to a set ratio, obtaining low-resolution three-dimensional image information according to one of the light signals, and obtaining two-dimensional image information according to the other light signal; Three-dimensional reconstruction is performed based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain a cell tissue morphology image of the 3D cell tissue sample to be tested.

2. The cell tissue morphology microscopic imaging method according to claim 1, characterized in that: The method of performing three-dimensional reconstruction based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain a cell tissue morphology image of a 3D cell tissue sample to be tested comprises: Two models are constructed, the first model is a mathematical model for simulating the Fourier stacking process under a virtual aperture, and the first model takes a three-dimensional point cloud as input and takes a spectrum and a phase map under different apertures as output, and the second model is a mathematical model for simulating an optical sensor, the input of which is a three-dimensional point cloud and the output is a three-dimensional image; The obtained low-resolution three-dimensional image information is used to construct three-dimensional point clouds with different focal lengths, and used as the initial input of the two models; the three-dimensional point cloud is used as the variable to be optimized, and regularization terms and constraint terms that conform to prior knowledge are added, and the outputs of the two models are compared with the obtained low-resolution three-dimensional image information and two-dimensional image information, and the input three-dimensional point cloud is optimized, and the cell tissue morphology image of the 3D cell tissue sample to be tested is obtained according to the optimized three-dimensional point cloud.

3. The cell tissue morphology microscopic imaging method according to claim 1, characterized in that: The method using Fourier stack illumination beam is: Light sources at different positions are illuminated in a redundant and overlapping manner, and the diffracted light with high-frequency and low-frequency information is collected separately.

4. The imaging device of the cell tissue morphology microscopic imaging method according to any one of claims 1 to 3, characterized in that: The device comprises: A light source module, used for parallelly irradiating a 3D cell tissue sample to be tested using a Fourier stacked illumination beam; An acquisition module is used to collect the diffracted light generated by the 3D cell tissue sample to be tested, divide the diffracted light into two optical signals according to a set ratio, obtain low-resolution three-dimensional image information according to one optical signal, and obtain two-dimensional image information according to the other optical signal; The three-dimensional reconstruction module is used to perform three-dimensional reconstruction based on the obtained low-resolution three-dimensional image information and two-dimensional image information to obtain a cell tissue morphology image of the 3D cell tissue sample to be tested.

5. The imaging device according to claim 4, characterized in that The light source module includes a two-dimensional LED array and an eyepiece lens group; The two-dimensional LED array adopts Fourier stack illumination, and the LED light of the two-dimensional LED array passes through the eyepiece lens group and irradiates the 3D cell tissue sample to be tested in parallel.

6. The imaging device according to claim 5, characterized in that The light source control system lights up the LEDs at different positions in the two-dimensional LED array in a time-sharing manner in an overlapping and redundant manner, so that the collection module collects the diffracted light with high-frequency and low-frequency information separately.

7. The imaging device according to claim 5, characterized in that The eyepiece lens group is a single lens group, a multi-lens group or a compound lens group.

8. The imaging device according to claim 4, characterized in that The acquisition module includes a microscope objective lens, a beam splitting prism, a two-dimensional image acquisition module and an optical sensor; The microscope objective collects the diffracted light generated by the 3D cell tissue sample to be tested, and the diffracted light is incident on the beam splitter prism, which splits the light into two light signals according to a set ratio, one of which enters the optical sensor and the other enters the two-dimensional image acquisition module; The optical sensor acquires low-resolution three-dimensional image information according to the one optical signal, and the two-dimensional image acquisition module acquires two-dimensional image information according to the other optical signal.

9. The imaging device according to claim 8, characterized in that: The optical sensor includes a tube lens group No. 1, a microlens array and a camera No. 1; The optical signal is sequentially passed through tube lens group No. 1 and microlens array, and then collected by camera No.

1. The image collected by camera No. 1 is low-resolution three-dimensional image information.

10. The imaging device according to claim 8, characterized in that The two-dimensional image acquisition module includes a tube lens group No. 2 and a camera No. 2; The other optical signal is collected by camera No. 2 through tube lens group No. 2, and the image collected by camera No. 2 is two-dimensional image information.

Images