Cell three-dimensional imaging device and method based on optical fiber vortex optical tweezers

Abstract

The invention discloses a cell three-dimensional imaging device and method based on optical fiber vortex optical tweezers, and relates to the technical field of cell three-dimensional imaging. An optical fiber vortex optical tweezers module in the device comprises a capture light source, an optical fiber isolator, an optical fiber coupler, a power meter and an optical fiber focusing vortex light generator; the optical fiber focusing vortex light generator is composed of a combined optical fiber and a spiral wave zone plate prepared at one end of the combined optical fiber, and one end with the spiral wave zone plate extends into the sample pool; incident laser is modulated by the combined optical fiber and the spiral wave zone plate to form a tightly focused vortex light beam; the tightly-focused vortex light beam forms a light trap at a focal point and is used for capturing a single cell; the vortex light beam carries orbital angular momentum and is used for driving captured cells to rotate; the high-speed microscopic imaging module is used for collecting projections of the rotating cells at different angles; and the image processing module is used for constructing a three-dimensional tomographic image of the single cell according to the projection at different angles.

Description

Technical Field

[0001] The present application relates to the technical field of three-dimensional cell imaging, and in particular to a three-dimensional cell imaging device and method based on fiber vortex optical tweezers. Background Art

[0002] In the biomedical field, single-cell tomography is crucial for in-depth investigations of cellular microstructure and function. Currently, three-dimensional images of biological tissues can be reconstructed by physically sectioning them or fluorescently labeling them, a technique known as invasive imaging. However, due to the tiny size of cells, this invasive imaging method is difficult to apply to cells. Summary of the Invention

[0003] The purpose of this application is to provide a cell three-dimensional imaging device and method based on fiber vortex optical tweezers, which can achieve damage-free three-dimensional imaging of cells.

[0004] To achieve the above objectives, this application provides the following solutions:

[0005] In a first aspect, the present application provides a cell three-dimensional imaging device based on fiber vortex optical tweezers, comprising:

[0006] Fiber vortex optical tweezers module, high-speed microscopy module and image processing module;

[0007] The fiber vortex optical tweezers module includes a capture light source, a fiber isolator, a fiber coupler, a power meter and a fiber focused vortex light generator; the capture light source is used to transmit the capture laser to the fiber isolator and then incident on the fiber isolator; the fiber coupler is used to couple the incident laser, and transmit one path of the coupled laser to the fiber focused vortex light generator, and one path of the laser to the power meter; the fiber focused vortex light generator is composed of a combined optical fiber and a spiral wave zone plate prepared at one end of the combined optical fiber; the end of the fiber focused vortex light generator prepared with the spiral wave zone plate is used to extend into the sample pool; the laser incident on the fiber focused vortex light generator is modulated by the combined optical fiber and the spiral wave zone plate to form a tightly focused vortex light beam; the tightly focused vortex light beam forms a light trap at the focus for capturing a single cell; the vortex light beam carries orbital angular momentum for driving the captured cell to rotate;

[0008] The high-speed microscopic imaging module includes an illumination device, an objective lens, a filter, and a camera; the illumination device is arranged directly above the sample pool; the camera is used to capture projections of cells at different angles during rotation; the filter filters out captured laser light; and the objective lens is used to magnify biological cells.

[0009] The image processing module is used to construct a three-dimensional tomographic image of a single cell based on projections at different angles.

[0010] Optionally, the capture light source is used to emit laser light of a set wavelength.

[0011] Optionally, the spiral zone plate has a focal length of less than 60 μm and a diameter of 120 μm.

[0012] Optionally, the image processing module is used to perform image preprocessing on projections of a single cell at different angles to obtain preprocessed projections; arrange the preprocessed projections according to the rotation angle to construct a three-dimensional matrix; slice the three-dimensional matrix along the optical axis to obtain a sinusoidal envelope diagram of the projection of a single cell; based on a filtered back projection algorithm, back-project the sinusoidal envelope diagram of the projection of a single cell to obtain a cell slice image; and stack the cell slice images along the optical axis to obtain a three-dimensional tomographic image of a single cell.

[0013] Optionally, the laser power emitted by the capture light source is greater than 260 mW and less than 350 mW.

[0014] In a second aspect, the present application provides a method for three-dimensional cell imaging based on fiber vortex optical tweezers, comprising:

[0015] The sample pool to be tested is placed on the fiber vortex optical tweezers module of the fiber vortex optical tweezers assisted cell three-dimensional imaging device;

[0016] Insert one end of the fiber vortex optical tweezers module with the spiral zone plate into the sample cell to be tested;

[0017] Adjust the input power of the capture light source to the set power value;

[0018] Observe the position of cells in the sample pool to be tested through the objective lens;

[0019] When the cells are stably captured, the position of the combined optical fiber is adjusted to the imaging center of the high-speed microscopy imaging module;

[0020] A high-speed camera is used to capture several projections of individual cells at different angles;

[0021] The projections of all individual cells at different angles are input into the image processing module to obtain a three-dimensional tomographic image.

[0022] Optionally, before placing the sample pool to be tested at the fiber vortex optical tweezers module, the method further includes:

[0023] The washed cell solution to be tested is placed in a sample pool to obtain a sample pool to be tested.

[0024] According to the specific embodiments provided in this application, this application discloses the following technical effects:

[0025] The present application provides a cell three-dimensional imaging device and method based on fiber vortex optical tweezers. The core of the device is a fiber-focused vortex beam generator, which consists of a combined optical fiber and a spiral zone plate prepared at one end of the combined optical fiber. The end of the spiral zone plate extends into a sample pool containing cells. When the laser passes through the combined optical fiber and the spiral zone plate, it is modulated into a tightly focused vortex beam. This vortex beam forms a light trap near the focus, which can capture a single cell. The vortex beam carries orbital angular momentum, which can be transferred to the captured cell, causing the cell to rotate around the optical axis. Through a high-speed microscopic imaging module, the rotating cells can be photographed from multiple angles to obtain projections of the cells at different angles. Based on the projections at different angles, a three-dimensional tomographic image of a single cell is constructed. The three-dimensional cell imaging of the present application uses optical tweezers technology to capture rotating cells contactlessly to obtain projection information. There is no need for invasive imaging such as slicing or fluorescent labeling of the cells, and it can achieve damage-free three-dimensional imaging of the cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0027] Figure 1 Schematic diagram of a fiber vortex optical tweezers-assisted three-dimensional cell imaging device provided in one embodiment of the present application;

[0028] Figure 2 A schematic diagram of the process of a method for three-dimensional cell imaging assisted by fiber vortex optical tweezers provided in one embodiment of the present application;

[0029] Figure 3 Schematic diagram of the capture and rotational manipulation of red blood cells by fiber vortex optical tweezers provided in one embodiment of the present application;

[0030] Figure 4 A schematic diagram of cell contour extraction provided in one embodiment of the present application;

[0031] Figure 5 Schematic diagram of segmentation projection of red blood cells at different rotation angles provided in one embodiment of the present application;

[0032] Figure 6 A schematic diagram of the red blood cell tomography process provided in one embodiment of the present application;

[0033] Figure 7 A schematic diagram of a tomographic slice of red blood cells in the xy, xz, and yz planes provided in one embodiment of the present application. DETAILED DESCRIPTION

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

[0035] As a non-invasive optical imaging technology, optical back-projection imaging technology has demonstrated unique advantages and great potential in single-cell research. Based on the principle of back-projection, it can reconstruct high-resolution three-dimensional images of single cells by collecting and processing projection data from multiple angles, and achieve detailed analysis of the internal structure and components of cells. This technology does not require fluorescent labeling of cells, avoiding the interference of the labeling process on the physiological state of cells, and can achieve non-destructive detection of cells, more realistically reflecting the natural state of cells. Fiber vortex optical tweezers use a tightly focused vortex beam with a spiral phase structure. The orbital angular momentum it carries can interact with cells to achieve stable capture and rotational manipulation of cells with multiple degrees of freedom, thereby obtaining multi-angle projection information of cells and reconstructing the three-dimensional structure of cells. This manipulation method is non-contact and low-damage, and is particularly suitable for operating fragile biological cells.

[0036] The purpose of this application is to provide a cell three-dimensional imaging device and method based on fiber vortex optical tweezers, which can achieve damage-free three-dimensional imaging of cells.

[0037] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the present application is further described in detail below with reference to the accompanying drawings and specific implementation methods.

[0038] Example 1

[0039] like Figure 1 As shown, this embodiment provides a cell three-dimensional imaging device based on fiber vortex optical tweezers, comprising:

[0040] Fiber vortex optical tweezers module, high-speed microscopy module and image processing module;

[0041] The fiber vortex optical tweezers module includes a capture light source, a fiber isolator, a fiber coupler, a power meter and a fiber focused vortex light generator; the capture light source is used to transmit the capture laser to the fiber isolator and then incident on the fiber isolator; the fiber coupler is used to couple the incident laser, and transmit one path of the coupled laser to the fiber focused vortex light generator, and one path of the laser to the power meter; the fiber focused vortex light generator is composed of a combined optical fiber and a spiral wave zone plate prepared at one end of the combined optical fiber; the end of the fiber focused vortex light generator prepared with the spiral wave zone plate is used to extend into the sample pool; the laser incident on the fiber focused vortex light generator is modulated by the combined optical fiber and the spiral wave zone plate to form a tightly focused vortex light beam; the tightly focused vortex light beam forms a light trap at the focus for capturing a single cell; the vortex light beam carries orbital angular momentum for driving the captured cell to rotate;

[0042] The high-speed microscopic imaging module includes an illumination device, an objective lens, a filter, and a camera; the illumination device is arranged directly above the sample pool; the camera is used to capture projections of cells at different angles during rotation; the filter filters out captured laser light; and the objective lens is used to magnify biological cells.

[0043] The image processing module is used to construct a three-dimensional tomographic image of a single cell based on projections at different angles.

[0044] This example builds on the optical angular momentum transfer properties of fiber-optic vortex tweezers to establish a multi-angle cell projection acquisition device, which then utilizes a back-projection algorithm to achieve 3D cell image tomography reconstruction. A fiber-optic tightly focused vortex beam generator is used to stably capture and rotate cells in three dimensions. High-speed image capture technology is combined to capture projection images of cells at different rotation angles. A 3D projection matrix is constructed sequentially according to the rotation angles to obtain a series of 2D projection slices of the cells. 3D tomographic imaging of single cells is then achieved using a filtered back-projection algorithm.

[0045] In the fiber vortex tweezers module, the captured light source exits through a fiber isolator and then connects to a fiber coupler, where it is split into two. One path is connected to a fiber focused vortex light generator, where it is regulated to form a tightly focused vortex beam at one end of a spiral zone plate. The other path is connected to a power meter to monitor the power in real time, thereby converting the input power to the vortex light generator based on the coupler ratio. A fiber focused vortex light generator can be implemented by fabricating a spiral zone plate on the end face of an optical fiber using focused ion beam or two-photon polymerization technology.

[0046] The combined optical fiber is composed of single-mode optical fiber and coreless optical fiber.

[0047] The high-speed microscopy module consists of an illumination light source, a filter, and a high-speed camera. The illumination light is used to illuminate the cells for imaging, while the filter is used to filter out scattered light from the capture laser beam hitting the cells. The high-speed camera features a high frame rate, enabling high-speed capture of projections of rotating cells.

[0048] Specifically, one end of the fiber focused vortex light generator, which is provided with a spiral zone plate, is used to extend into the sample pool, which is a solution pool containing cells on the zone plate.

[0049] Among them, the image processing module is used to perform image preprocessing on the projections of a single cell at different angles to obtain preprocessed projections; the preprocessed projections are arranged according to the rotation angle to construct a three-dimensional matrix; the three-dimensional matrix is sliced along the optical axis to obtain a sinusoidal envelope diagram of the projection of a single cell; based on the filtered back projection algorithm, the sinusoidal envelope diagram of the projection of a single cell is back-projected to obtain a cell slice image; and the cell slice images are stacked along the optical axis to obtain a three-dimensional tomographic image of a single cell.

[0050] Example 2

[0051] This embodiment provides a method for three-dimensional cell imaging based on fiber vortex optical tweezers, comprising:

[0052] The sample pool to be tested is placed on the fiber vortex optical tweezers module of the fiber vortex optical tweezers assisted cell three-dimensional imaging device;

[0053] Insert one end of the fiber vortex optical tweezers module with the spiral zone plate into the sample cell to be tested;

[0054] Adjust the input power of the capture light source to the set power value;

[0055] Observe the position of cells in the sample pool to be tested through the objective lens;

[0056] After the red blood cells are stably captured, the position of the combined optical fiber is adjusted to the imaging center of the high-speed microscopy imaging module;

[0057] A high-speed camera is used to capture several projections of individual cells at different angles;

[0058] The projections of all individual cells at different angles are input into the image processing module to obtain a three-dimensional tomographic image.

[0059] Before placing the sample pool to be tested on the fiber vortex optical tweezers module, the method further includes:

[0060] The washed cell solution to be tested is placed in a sample pool to obtain a sample pool to be tested.

[0061] Specifically, the three-dimensional cell imaging method in this embodiment can be as follows: Figure 2 As shown,

[0062] 1) Cell capture and rotation manipulation:

[0063] By adding the cell solution to the sample pool on the wave plate, turning on the capture laser and adjusting the output power, stable capture of the cells and rotation around the optical axis can be achieved, while ensuring that the laser power is not too high to damage the cells.

[0064] 2) Multi-angle projection acquisition:

[0065] Adjust the position of the objective lens so that the camera presents a clear image of the cell. Use the acquisition card to collect a series of projections during the cell rotation process, and determine the corresponding rotation angle of the cell based on the cell projection characteristics.

[0066] 3) Cell 3D tomographic image reconstruction:

[0067] All projections of a group of cells within a 180° rotation are selected and background subtracted and noise filtered. The projections are then arranged according to the rotation angle to construct a three-dimensional matrix. The three-dimensional matrix is sliced along the optical axis to obtain a sinusoidal envelope of the cell projections. Cell slice images are then calculated through backprojection, and finally stacked along the optical axis to form a three-dimensional tomographic image of the cells.

[0068] This application also provides a specific implementation case:

[0069] The capture and rotation of red blood cells are achieved based on the fiber vortex optical tweezers system, which enables three-dimensional tomographic imaging of red blood cells. The details are as follows:

[0070] Fiber vortex optical tweezers module in the implementation case:

[0071] The capture light source is a 1064nm laser. The absorption coefficient of near-infrared laser in liquid water environment is low, which greatly reduces the thermal damage caused by the absorption of laser energy by water molecules, ensuring the activity of living cells during the capture process. At the same time, the near-infrared light has no overlap with the resonance absorption peak (ultraviolet-visible light region) of chromophores (such as proteins and nucleic acids) in the cells, reducing the interference of photochemical reactions on cells. In addition, 1064nm is a low-loss window for fiber optic communication (especially in single-mode optical fiber), which is suitable for long-distance transmission and high-quality focused vortex beam generation of fiber vortex optical tweezers systems. The fundamental mode light wavelength corresponding to the fiber isolator, fiber coupler, and power meter is 1064nm.

[0072] The fiber-focused vortex light generator uses a focused ion beam to etch a spiral zone plate on the end face of an optical fiber. The zone plate has a focal length of 60 μm and a diameter range of 120 μm. With these parameters, the zone plate can transform the Gaussian base mode beam in the fiber into a focused vortex beam with a numerical aperture greater than 0.8, sufficient for three-dimensional capture of red blood cells. Red blood cells are 6-8 μm in size, and the topological charge of the zone plate is 3, generating a vortex beam with a focused spot diameter smaller than that of the red blood cell. This vortex beam then induces spin in the red blood cell as it transfers angular momentum to it.

[0073] The illumination is LED white light, and the filter uses a 600nm wavelength low-pass filter to filter scattered light from the captured laser. The high-speed camera achieves a full-frame frame rate of 1280 x 1024 at 1050 fps, and a maximum frame rate of 7500 fps with reduced resolution and windowing. The pixel size is 6.6μm x 6.6μm, and the shutter speed is 100μs. It also features 1TB of dedicated high-speed image data storage and a CoaxPress data transmission interface with a transmission bandwidth of at least 2GB / s. The high frame rate ensures that a sufficient number of images from different angles can be captured during cell rotation, while the high resolution ensures image clarity for accurate analysis of cell morphological features. The fast shutter speed effectively reduces motion blur, ensuring clear projection images during cell rotation. The camera's fast data transmission speed enables timely transfer of large amounts of captured image data to a computer for storage and subsequent processing.

[0074] Specific implementation steps for 3D imaging of red blood cells:

[0075] 1. Red blood cell capture and rotation manipulation:

[0076] A cleaned red blood cell solution is placed in a sample cell, and an optical fiber is extended horizontally into the sample cell. The surface tension of the liquid allows the fiber endface to be immersed in the red blood cell solution. By adjusting the imaging plane of the objective lens and the position of the optical fiber, a clear image of the fiber endface is formed within the field of view. The capture laser input power is adjusted to 300mW. When the focused vortex light is applied to the red blood cells, the radiation pressure and gradient forces gradually attract the red blood cells to the center of the light field, which is the capture area of the optical tweezers. During the capture process, the position and state of the red blood cells are observed in real time using a microscope. Once the red blood cells are stably captured, the position of the optical fiber is adjusted to manipulate the red blood cells to the center of the image. High-power lasers cause red blood cells to absorb excessive energy, causing a sharp increase in the cell's internal temperature. Biomacromolecules such as proteins within red blood cells are very sensitive to temperature. Excessive temperatures can denature proteins and disrupt the structure and function of the cell membrane. Once the integrity of the cell membrane is compromised, intracellular substances leak out, causing red blood cell lysis. During the capture process, the input power should be ensured to be lower than the tolerance power threshold of the red blood cells and at the same time be able to drive the red blood cells to rotate uniformly. In the experimental case used, the input laser power should be greater than 260mW and less than 350mW.

[0077] 2. Multi-angle projection acquisition of red blood cells:

[0078] A series of images of red blood cell rotation were collected by a high-speed camera, such as Figure 3 The conventional method is to find the cell's characteristic points on the red blood cell, track their positions in each frame using a computer, and then calculate the corresponding rotation angle information. Red blood cells have a distinct pancake-like morphology, so during rotation, the thickness of their projection changes with the angle. Rotation is observed by extracting the width change of the red blood cell projection at the center of the image, and the rotation angle corresponding to each frame is determined based on the width change.

[0079] 3. Reconstruction of 3D tomographic images of red blood cells:

[0080] Select the projection frames of the acquired red blood cells within the range of 180°, perform Gaussian filtering on each frame of the projection image to remove Gaussian noise, smooth the image, reduce the impact of noise on subsequent segmentation, and improve image quality. Use methods such as histogram equalization and adaptive histogram equalization to expand the grayscale differences between cells and background, and between cells in the image, make the boundaries of cells clearer, highlight the characteristics of cells, and detect the edges of cells. Subsequently, perform morphological operations such as corrosion, expansion, opening, and closing operations. Corrosion can remove small noise and burrs in the image, expansion can fill small holes inside the cells, and opening and closing operations are used to eliminate small protrusions and small depressions in the image, respectively, to further optimize the outline of the cells and highlight the morphological characteristics of the cells, such as Figure 4As shown. Finally, the cell boundary is extracted, and morphological operations are applied again to further optimize the segmentation results, remove the small noise areas remaining in the segmented image, fill small holes, etc., so that the segmented cell area is more complete and accurate. In order to reduce the error caused by jitter, the projection is calibrated for position center and rotation axis. Based on the center calibration algorithm of centroid calculation, for each cell segmentation projection image, the centroid of the red blood cell area is calculated, and the centroid is used as the center position of the red blood cell in the projection image. Then, the red blood cell centers in all projection images are adjusted to the same position through translation transformation to obtain the projection sequence after center calibration. A rotation axis calibration algorithm based on feature point matching is adopted. First, feature points are selected in the projection image after center calibration. These feature points should have obvious features and be relatively stable during the rotation of red blood cells. Then, in subsequent projection images, the corresponding positions of these feature points at different angles are found through image matching algorithm. According to the displacement information of the feature points and the known red blood cell rotation angle, the offset of the rotation axis can be calculated. By correcting the rotation axis, the rotation axis of all projection images is kept consistent, and the projection sequence after rotation axis calibration is obtained. Figure 5 The calibrated cell segmentation projections at 0°, 45°, 90°, 135°, and 180° are shown. The segmented cell projections are arranged according to the rotation angle to form a three-dimensional matrix, as shown in FIG. Figure 6 As shown, the matrix is sliced laterally and then Fourier transformed to obtain a sine envelope sequence. All sine envelopes are back-projected to obtain axial slices of the cell. The slices are stacked axially to form a three-dimensional image of the cell, as shown in FIG. Figure 7 The three-dimensional tomographic images are shown as xy, xz and yz.

[0081] In summary, this application has the following technical effects:

[0082] (1) The three-dimensional cell imaging described in this application uses optical tweezers technology to capture rotating cells without contact to obtain projection information. There is no need for invasive imaging such as slicing or fluorescent labeling of cells, and it can achieve non-destructive three-dimensional imaging of cells.

[0083] (2) The fiber optic vortex tweezers described in this application assist in three-dimensional cell imaging. The fiber optics have a flexible manipulation space and the ability to invasively manipulate cells in vivo. This makes this imaging method promising for invasive cell imaging in vivo.

[0084] (3) The spot size of the fiber vortex tweezers can be controlled by the topological charge of the vortex beam, which can capture and rotate cells of different sizes, thereby achieving different three-dimensional imaging of cells. By selecting illumination light of different wavelengths, it is possible to perform tomographic imaging of cells of different transparencies. For example, selecting illumination light of the infrared band has stronger penetrating power, so it is possible to perform tomographic imaging of more complex cells.

[0085] The technical features of the above embodiments can be combined arbitrarily. To make the description concise, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0086] This document uses specific examples to illustrate the principles and implementation methods of this application. The description of the above examples is only intended to help understand the method and core concept of this application. At the same time, for those skilled in the art, based on the concept of this application, there may be changes in the specific implementation methods and application scope. In summary, the content of this specification should not be understood as limiting this application.

Claims

1. A cell three-dimensional imaging device based on fiber vortex optical tweezers, characterized in that: include: Fiber vortex optical tweezers module, high-speed microscopy imaging module and image processing module; The fiber vortex optical tweezers module includes a capture light source, a fiber isolator, a fiber coupler, a power meter and a fiber focused vortex light generator; the capture light source is used to transmit the capture laser to the fiber isolator and then incident on the fiber isolator; the fiber coupler is used to couple the incident laser, and transmit one path of the coupled laser to the fiber focused vortex light generator, and one path of the laser to the power meter; the fiber focused vortex light generator is composed of a combined optical fiber and a spiral wave zone plate prepared at one end of the combined optical fiber; the end of the fiber focused vortex light generator prepared with the spiral wave zone plate is used to extend into the sample pool; the laser incident on the fiber focused vortex light generator is modulated by the combined optical fiber and the spiral wave zone plate to form a tightly focused vortex light beam; the tightly focused vortex light beam forms a light trap at the focus for capturing a single cell; the vortex light beam carries orbital angular momentum for driving the captured cell to rotate; The high-speed microscopic imaging module includes an illumination device, an objective lens, a filter, and a camera; the illumination device is arranged directly above the sample pool; the camera is used to capture projections of cells at different angles during rotation; the filter filters out captured laser light; and the objective lens is used to magnify biological cells. The image processing module is used to construct a three-dimensional tomographic image of a single cell based on projections at different angles.

2. A cell three-dimensional imaging device based on fiber vortex optical tweezers according to claim 1, characterized in that, The capture light source is used to emit laser light of a set wavelength.

3. A cell three-dimensional imaging device based on fiber vortex optical tweezers according to claim 1, characterized in that, The spiral zone plate has a focal length of less than 60 μm and a diameter of 120 μm.

4. A cell three-dimensional imaging device based on fiber vortex optical tweezers according to claim 1, characterized in that, The image processing module is used to perform image preprocessing on projections of individual cells at different angles to obtain preprocessed projections; the preprocessed projections are arranged according to the rotation angles to construct a three-dimensional matrix; The three-dimensional matrix is sliced along the optical axis to obtain the sinusoidal envelope of the projection of a single cell; Based on the filtered back-projection algorithm, the sinusoidal envelope of the projection of a single cell is back-projected to obtain a cell slice image; the cell slice images are stacked along the optical axis to obtain a three-dimensional tomographic image of a single cell.

5. A cell three-dimensional imaging device based on fiber vortex optical tweezers according to claim 1, characterized in that, The laser power emitted by the capture light source is greater than 260 mW and less than 350 mW.

6. A method for three-dimensional cell imaging based on fiber vortex optical tweezers, characterized in that: include: Placing the sample pool to be tested at the fiber vortex optical tweezers module of the fiber vortex optical tweezers assisted cell three-dimensional imaging device according to any one of claims 1 to 5; Insert one end of the fiber vortex optical tweezers module with the spiral zone plate into the sample cell to be tested; Adjust the input power of the capture light source to the set power value; Observe the position of cells in the sample pool to be tested through the objective lens; When the cells are stably captured, the position of the combined optical fiber is adjusted to the imaging center of the high-speed microscopy imaging module; A high-speed camera is used to capture several projections of individual cells at different angles; The projections of all individual cells at different angles are input into the image processing module to obtain a three-dimensional tomographic image.

7. A method for three-dimensional cell imaging based on fiber vortex optical tweezers according to claim 6, characterized in that: Before placing the sample pool to be tested on the fiber vortex optical tweezers module, the following steps are also included: The washed cell solution to be tested is placed in a sample pool to obtain a sample pool to be tested.

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