Three-dimensional fluorescence imaging method, device, equipment and medium based on non-diffracting beam

Abstract

The present application relates to the field of optical imaging technology, and discloses a three-dimensional fluorescence imaging method and device based on non-diffracting light beams, electronic equipment and medium, the method comprising: collecting a first fluorescence projection image and a second fluorescence projection image of a preset sample, and extracting axial projection information of the sample according to the first fluorescence projection image; extracting lateral displacement offset information of the sample according to the second fluorescence projection image; signal pairing the first fluorescence projection image and the second fluorescence projection image according to the axial projection information and the lateral displacement offset information to obtain a fluorescence signal set; and stacking the fluorescence signal set into a three-dimensional fluorescence image corresponding to the sample according to a preset stacking rule. By extracting the axial projection information and the lateral displacement offset information, image signal pairing is completed, signal redundancy and matching errors caused by traditional scanning are avoided, signal screening efficiency is improved, and fast high-resolution three-dimensional imaging is realized.

Description

Technical Field

[0001] This invention relates to the field of optical imaging technology, and in particular to a three-dimensional fluorescence imaging method, apparatus, device, and medium based on a diffraction-free beam. Background Technology

[0002] In fields such as biomedical testing, material microstructure analysis, and precision optical measurement, obtaining the three-dimensional spatial distribution information of fluorescent samples is a core technical requirement. For example, scenarios such as cell localization of fluorescent labels in biological tissues, distribution detection of fluorescent microspheres inside materials, and three-dimensional morphology characterization of micro-nano devices all require accurate acquisition of the position, morphology, and intensity information of fluorescent substances in three-dimensional space.

[0003] Currently, traditional fluorescence imaging methods typically reconstruct the three-dimensional distribution information of samples by scanning layer by layer along the axis. However, this method requires multiple scans of the sample, which can lead to problems such as limited imaging speed and high system complexity, making it difficult to meet the needs of high temporal resolution volume imaging of rapid biological processes.

[0004] Therefore, in the face of the growing demand for fluorescence imaging, current fluorescence imaging methods urgently need to be improved to solve the problem of low fluorescence imaging efficiency of existing methods. Summary of the Invention

[0005] This invention provides a method, apparatus, device, and medium for three-dimensional fluorescence imaging based on a non-diffraction beam, the main purpose of which is to solve the problem of low efficiency in three-dimensional fluorescence imaging based on a non-diffraction beam.

[0006] To achieve the above objectives, the present invention provides a three-dimensional fluorescence imaging method based on a diffraction-free beam, comprising:

[0007] Acquire the first fluorescence projection image and the second fluorescence projection image of the preset sample, and extract the axial projection information of the sample based on the first fluorescence projection image;

[0008] The lateral displacement information of the sample is extracted based on the second fluorescence projection image;

[0009] Based on the axial projection information and the lateral displacement offset information, the first fluorescence projection image and the second fluorescence projection image are paired to obtain a set of fluorescence signals;

[0010] The fluorescence signals are stacked into a three-dimensional fluorescence image corresponding to the sample according to the preset stacking rules.

[0011] The present invention also provides a three-dimensional fluorescence imaging device based on a diffraction-free beam, the device comprising:

[0012] The axial projection information extraction module is used to acquire the first fluorescence projection image and the second fluorescence projection image of the preset sample, and extract the axial projection information of the sample based on the first fluorescence projection image.

[0013] The lateral displacement information extraction module is used to extract the lateral displacement information of the sample based on the second fluorescence projection image.

[0014] The fluorescence information set pairing module is used to pair the first fluorescence projection image with the second fluorescence projection image based on axial projection information and lateral displacement offset information to obtain a fluorescence signal set.

[0015] The fluorescence image stacking module is used to stack the fluorescence signal sets into a three-dimensional fluorescence image corresponding to the sample according to a preset stacking rule.

[0016] The present invention also provides an electronic device, the electronic device comprising:

[0017] At least one processor; and,

[0018] A memory that is communicatively connected to at least one processor; wherein,

[0019] The memory stores a computer program that can be executed by at least one processor, which enables the at least one processor to perform the above-described three-dimensional fluorescence imaging method based on a diffraction-free beam.

[0020] The present invention also provides a computer-readable storage medium storing at least one computer program, which is executed by a processor in an electronic device to implement the above-described three-dimensional fluorescence imaging method based on a diffraction-free beam.

[0021] This invention, through the acquisition of dual fluorescence projection images and extraction of axial projection information, eliminates the need for layer-by-layer axial scanning. A single acquisition of dual projection images is sufficient to obtain the axial attributes of the fluorescence signal, avoiding the multiple imaging operations required by traditional layer-by-layer scanning and significantly reducing data acquisition time. By extracting lateral displacement information, a dual matching dimension of axial and lateral dimensions is formed, enriching feature data without requiring additional scans and further improving subsequent pairing efficiency. Image signal pairing is completed using axial and lateral information, avoiding signal redundancy and matching errors caused by multiple scans in traditional methods, thus improving signal filtering efficiency. By stacking fluorescence signal sets according to rules, the traditional layer-by-layer image stitching is eliminated, thereby reducing system control complexity and achieving rapid, high-resolution three-dimensional imaging. Attached Figure Description

[0022] Figure 1 This is a schematic flowchart of a three-dimensional fluorescence imaging method based on a diffraction-free beam provided in an embodiment of the present invention.

[0023] Figure 2 A functional block diagram of a three-dimensional fluorescence imaging system based on a diffraction-free beam provided in an embodiment of the present invention;

[0024] Figure 3 An axial intensity distribution diagram and corresponding phase pattern of a non-diffractive beam are provided in an embodiment of the present invention.

[0025] Figure 4 A functional block diagram of a three-dimensional fluorescence imaging device based on a diffraction-free beam provided in an embodiment of the present invention;

[0026] Figure 5 This is a schematic diagram of an electronic device for implementing a three-dimensional fluorescence imaging method based on a diffraction-free beam, according to an embodiment of the present invention.

[0027] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0028] It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0029] To address the limitations of existing three-dimensional fluorescence imaging methods based on diffraction-free beams, which require multiple scans of the sample and suffer from limitations in imaging speed and high system complexity, making it difficult to meet the demand for high temporal resolution volumetric imaging of rapid biological processes, this application provides a three-dimensional fluorescence imaging method based on diffraction-free beams. This method extracts axial projection information and lateral displacement information to complete image signal pairing, avoiding signal redundancy and matching errors caused by traditional scanning, improving signal screening efficiency, and thus achieving rapid high-resolution three-dimensional imaging.

[0030] Reference Figure 1 The diagram shown is a flowchart illustrating a three-dimensional fluorescence imaging method based on a diffraction-free beam according to an embodiment of this application. In this embodiment, the three-dimensional fluorescence imaging method based on a diffraction-free beam includes:

[0031] S1. Acquire the first fluorescence projection image and the second fluorescence projection image of the preset sample, and extract the axial projection information of the sample based on the first fluorescence projection image.

[0032] In this embodiment of the invention, the preset sample can be an agarose sample with a certain thickness and encapsulated with fluorescent microspheres. The fluorescence projection image includes a first fluorescence projection image and a second fluorescence projection image, which can be a two-dimensional fluorescence image acquired after illuminating the same area of ​​the preset sample with a phase-map modulated non-diffraction beam (including the first non-diffraction beam and the second non-diffraction beam).

[0033] In this embodiment of the invention, axial projection information refers to the feature data extracted from the first fluorescence projection image acquired after the sample is illuminated by the first non-diffractive beam. It includes attributes such as the intensity and grayscale distribution of the fluorescence signal in the sample after superimposing the projection along the optical axis, and also includes the two-dimensional coordinate information of the fluorescence signal region in the horizontal plane.

[0034] like Figure 2 As shown, the three-dimensional fluorescence imaging system 200 based on a non-diffraction beam includes a light source module 201, a beam shaping module 202, a scanning module 203, a sample illumination module 204, and an imaging module 205.

[0035] Furthermore, during the acquisition of the first fluorescence projection image of the preset sample, a Gaussian beam is first excited using the light source module 201. The Gaussian beam is incident on the beam shaping module 202, which includes a Fourier lens and a high-pass filter. That is, the Gaussian beam illuminates the front focal plane of the Fourier lens, and a high-pass filter is placed on the front focal plane of the Fourier lens. The high-pass filter blocks the low-frequency light field passing through the Gaussian beam, allowing only the high-frequency light field of the Gaussian beam to pass through. Subsequently, a first non-diffraction beam is formed at the rear focal plane of the Fourier lens. The first non-diffraction beam is deflected and controlled by the scanning module 203. The sample illumination module 204 illuminates the preset area of ​​the sample with the controlled first non-diffraction beam, exciting the fluorescent material in the preset area of ​​the sample to generate a corresponding fluorescence signal. The fluorescence signal is then captured by the imaging module 205 and converted into image data to obtain the first fluorescence projection image. The first non-diffraction beam can be a Bessel beam, an approximate Bessel beam, or an axisymmetric beam with extended depth of focus.

[0036] In this embodiment of the invention, the transmittance of the high-pass filter is expressed by the following formula:

[0037]

[0038] in, This represents the transmittance of the high-pass filter; when the transmittance of the high-pass filter is 1, light in the corresponding area can pass through completely; when the transmittance of the high-pass filter is 0, light in the corresponding area is completely blocked. Let be the inner ring radius of the high-pass filter. This is the effective aperture of the high-pass filter.

[0039] Furthermore, during the acquisition of the second fluorescence projection image of the preset sample, a Gaussian beam is also excited using the light source module 201. The Gaussian beam is incident on the beam shaping module 202, which also includes a spatial light modulator and a Fourier lens. The Gaussian beam illuminates the modulation plane of the spatial light modulator, where a specific phase distribution map calculated according to a preset bending trajectory is loaded (the specific phase distribution map is composed of phase distribution data, which are quantized phase values; the specific phase distribution map is a visual representation of the phase distribution data). After the Gaussian beam is modulated by the specific phase distribution map, the modulated beam passes through a Fourier lens. The light field distribution of the beam is formed on the back focal plane; finally, the light field distribution of the beam is used as the second non-diffractive beam, and the second non-diffractive beam is deflected by the scanning module 203 so that the illuminated area of ​​the deflected second non-diffractive beam is consistent with the sample illumination area of ​​the first non-diffractive beam when it is illuminated; then the sample illumination module 204 illuminates the area of ​​the sample illuminated by the first non-diffractive beam with the deflected second non-diffractive beam, exciting the fluorescent material in the preset area of ​​the sample to generate a corresponding fluorescence signal, and then the imaging module 205 captures the fluorescence signal and converts it into image data, thereby obtaining the second fluorescence projection image, wherein the second non-diffractive beam is a beam that presents a parabolic or curved propagation trajectory during propagation.

[0040] The phase distribution data is expressed by the following formula:

[0041]

[0042] in, This refers to the phase distribution data loaded onto the spatial light modulator. This indicates a phase take operation. Indicates the number of focal points in the focused area. Indicates the first A standard cubic phase, Indicates the first The light intensity ratio parameter of each focal point Indicates the first The horizontal coordinates of the focal point in the space of the back focal plane of the Fourier lens. Indicates the first The vertical coordinates of each focal point in the space of the back focal plane of the Fourier lens. Indicates the first The coordinates of each focal point along the optical axis (propagation direction) in the space of the back focal plane of the Fourier lens. Indicates the first Initial phase shift of each focus, This refers to the phase function; where, and The values ​​must be consistent.

[0043] The standard cubic phase is expressed by the following formula:

[0044]

[0045] in, Indicates the first A standard cubic phase, Represents a complex exponential function. Represents the imaginary unit. Indicates the first The cubic phase period of each focal point is used to determine the spatial repetition frequency of the phase pattern. The larger the value, the faster the phase change and the higher the lateral deflection rate of the beam. Represents the wavenumber of light. Indicates the first The converging angle of a focal point Indicates the first Control parameters for the cubic phase distribution of each focus. Indicates the maximum convergence angle of the objective lens. Indicates the first The azimuth of each focal point, Indicates the first The initial rotation angle of each focus; where, and The values ​​must be consistent.

[0046] In this embodiment of the invention, extracting axial projection information of a sample from a first fluorescence projection image includes: identifying a fluorescence signal region in the first fluorescence projection image corresponding to the sample; performing grayscale distribution analysis on the fluorescence signal region and extracting axial projection grayscale features of the fluorescence signal region based on the analysis results; calibrating the signal intensity of the axial projection grayscale features according to preset acquisition parameters to obtain calibrated axial features; and associating the calibrated axial features with the corresponding fluorescence signal region to generate axial projection information corresponding to the fluorescence signal region.

[0047] In this embodiment of the invention, in the process of identifying the fluorescence signal region corresponding to the sample in the first fluorescence projection image, the first fluorescence projection image can first be preprocessed using a Gaussian filtering algorithm. The preprocessing includes noise reduction and image enhancement to obtain the preprocessed first fluorescence projection image. Then, a grayscale threshold is set, and pixels with grayscale values ​​greater than the grayscale threshold are selected from the preprocessed first fluorescence projection image and determined as sample fluorescence signal pixels. Morphological closing operations are performed on the determined pixels to obtain the fluorescence signal region corresponding to the sample.

[0048] Furthermore, in the process of analyzing the gray-level distribution of the fluorescence signal region, the number of pixels at each gray level in the fluorescence signal region is first counted to obtain the gray-level histogram of the region. Then, features such as the shape, peak position, and distribution range of the gray-level histogram are extracted. Based on the extracted features, the gray-level distribution in the fluorescence signal region is analyzed, which is the analysis result. From the analysis result, information that can reflect the axial projection gray-level characteristics of the fluorescence signal region, such as gray-level mean and gray-level variance, is extracted.

[0049] Furthermore, during the signal intensity calibration of the axial projection grayscale features, the preset acquisition parameters include excitation power, exposure time, detector gain, and dark noise. Based on the changes in the preset acquisition parameters, the axial projection grayscale features are scaled or offset accordingly, so that the calibrated axial features can more accurately reflect the actual fluorescence signal intensity of the sample and eliminate the influence of changes in acquisition parameters. For each pixel in the fluorescence signal region, the calibrated axial feature corresponding to the pixel is associated with the coordinates of the pixel to generate the axial projection information corresponding to the fluorescence signal region.

[0050] For example, changes in acquisition parameters cause proportional changes in the signal. The axial projection grayscale features are multiplied by a scaling factor (such as multiplying by 2.0) to compensate for the attenuation of signal strength.

[0051] In this embodiment of the invention, the intensity of the acquired parameter calibration signal is used to eliminate the influence of parameter changes, so that the calibration axial features more accurately reflect the sample fluorescence signal; the calibration features are associated with the regional coordinates to generate axial projection information, which can intuitively present the positional distribution of the sample fluorescence signal.

[0052] like Figure 3 As shown, this represents two sets of non-diffractive beam combinations with different axial lengths: the first set is (b) and (d), with an axial length of 10 μm, where (b) is the numerical aperture of the objective lens. The first non-diffractive beam at time (a) is the corresponding phase pattern; (d) represents... , , , and The second non-diffractive beam at that time, (c) is the corresponding phase pattern; the second set is (f) and (h), with an axial length of 20 μm, where (f) represents the phase pattern. The first non-diffractive beam at time (e) represents the corresponding phase pattern; while (h) represents... , , , and The second non-diffractive beam at time (g) is the corresponding phase pattern.

[0053] S2. Extract the lateral displacement information of the sample based on the second fluorescence projection image.

[0054] In this embodiment of the invention, lateral displacement offset information refers to quantitative data that reflects the systematic positional shift of fluorescence signals at different depths (axial positions) within the sample on the imaging plane; it assists in the analysis of dynamic characteristics such as sample motion trajectory, interaction, and structural changes.

[0055] In this embodiment of the invention, extracting the lateral displacement offset information of the sample based on the second fluorescence projection image includes: identifying a homologous fluorescence signal region in the second fluorescence projection image corresponding to the fluorescence signal region; determining the second lateral coordinate of the fluorescence signal region in the second fluorescence projection image based on the homologous fluorescence signal region; extracting the first lateral coordinate of the fluorescence signal region in the first fluorescence projection image; calculating the difference between the first lateral coordinate and the second lateral coordinate, and using the difference as the lateral displacement offset of the fluorescence signal region; and calculating the lateral displacement offset information corresponding to the fluorescence signal region using a preset displacement-axial position mapping formula based on a preset lateral coordinate constant and the lateral displacement offset.

[0056] In this embodiment of the invention, during the process of identifying the same source fluorescence signal region, the unique visual features of the fluorescence signal region are first extracted, such as brightness distribution pattern, edge contour shape or texture details. Then, image features consistent with the unique visual features are extracted in the second fluorescence projection image. The unique visual features and image features are compared for similarity. Based on the similarity of the compared features (such as shape matching degree, brightness distribution consistency), the corresponding region of the fluorescence signal region in the second fluorescence projection image (i.e., the same source fluorescence signal region) is determined. The coordinate position of the corresponding region in the second fluorescence projection image is determined, and the coordinate position is used as the second horizontal coordinate.

[0057] In this embodiment of the invention, the step of extracting the first horizontal coordinate of the fluorescence signal region in the first fluorescence projection image is the same as the step of determining the second horizontal coordinate of the fluorescence signal region in the second fluorescence projection image based on the same fluorescence signal region, and will not be described in detail here.

[0058] Furthermore, by comparing the positional differences between the first and second lateral coordinates in the horizontal direction of the image, the difference between the two is calculated. This difference reflects the relative change in the lateral position of the fluorescence signal region during the two imaging processes. For example, if the first lateral coordinate is above and the second lateral coordinate is below, the difference is positive, indicating an upward shift; otherwise, it is a downward shift.

[0059] Furthermore, the displacement-axial position mapping formula can be expressed as:

[0060]

[0061] in, This refers to the first horizontal coordinate. This refers to the second horizontal coordinate. This refers to the absolute value of the difference. It refers to any constant x-axis value, with units of 1. , The coordinates of the axial position of the fluorescence signal region are given. This refers to the magnitude of the wave vector;

[0062] The axial position coordinates corresponding to the lateral displacement offset are obtained from the formula. The lateral displacement offset and the axial position coordinates are mapped to form a lateral displacement offset-axial position mapping table, which is the lateral displacement offset information.

[0063] In this embodiment of the invention, the difference between the first horizontal coordinate and the second horizontal coordinate is calculated to eliminate interference from different signal regions, providing accurate basic data for the subsequent generation of effective offset information. By using a mapping formula to associate the horizontal displacement offset with the horizontal coordinate constant, the limitation of two-dimensional plane dimension is broken, providing a reliable quantitative basis for the accurate pairing of subsequent fluorescence signals and high-quality three-dimensional fluorescence image reconstruction.

[0064] S3. Based on the axial projection information and the lateral displacement offset information, the first fluorescence projection image and the second fluorescence projection image are paired to obtain a set of fluorescence signals.

[0065] In this embodiment of the invention, the fluorescence signal set refers to the set of paired results used to characterize the correspondence between signal regions belonging to the same physical luminescent point in the first and second fluorescence projection images. This provides verified and accurate basic data on the correspondence of signal points for subsequent recovery of the three-dimensional spatial distribution information of the sample from the two-dimensional projection image.

[0066] In this embodiment of the invention, a first fluorescence projection image and a second fluorescence projection image are paired based on axial projection information and lateral displacement offset information to obtain a fluorescence signal set. This includes: extracting spatial distribution features of the fluorescence signal region from the axial projection information and extracting lateral displacement features of the fluorescence signal region from the lateral displacement offset information; converting the spatial distribution features and lateral displacement features into spatial distribution feature vectors and lateral displacement vectors respectively; determining the region feature descriptions corresponding to the fluorescence signal regions based on the spatial distribution feature vectors, lateral displacement vectors, and first and second lateral coordinates; calculating the feature similarity of the fluorescence signal regions in the first and second fluorescence projection images based on the region feature descriptions; matching the fluorescence signal regions in the first fluorescence projection image with the fluorescence signal regions in the second fluorescence projection image with the highest similarity based on the feature similarity to form a preliminary pairing result set; and performing multiple constraint checks on the preliminary pairing result set to obtain a fluorescence signal set that passes the checks.

[0067] In this embodiment of the invention, during the extraction of spatial distribution features, the image performance features of each fluorescence signal region are first analyzed based on the axial projection information. The image performance features include the brightness distribution concentration of the region, the morphological expansion trend on the two-dimensional plane (e.g., dot-like or with a certain elongation direction), and the intensity gradient change law from the center of the region to the edge. Then, the image performance features are converted into numerical indicators (e.g., the equivalent radius of the light spot, the second moment of intensity, etc.) to generate a set of image feature parameters, which are defined as spatial distribution features.

[0068] Furthermore, in the process of extracting lateral offset features, the inherent properties of the lateral displacement are extracted from the lateral displacement information corresponding to the fluorescence signal region. The inherent properties include: the absolute magnitude of the displacement (i.e., the size of the offset pixel distance), the dominant direction of the displacement (such as biased towards the positive or negative x-axis of the image coordinate system), etc., and the inherent properties are used as the lateral offset features corresponding to the fluorescence signal region.

[0069] Furthermore, the similarity calculation is performed between the regional feature descriptions corresponding to the fluorescence signal regions in the first fluorescence projection image and the regional feature descriptions corresponding to each fluorescence signal region in the second fluorescence projection image. The similarity calculation refers to determining whether the feature vectors (spatial distribution feature vector and lateral offset vector) of the two regions are highly consistent in terms of direction and other indicators, and whether the coordinates of the two regions are close within a reasonable spatial range.

[0070] Furthermore, by traversing all fluorescence signal regions of the second fluorescence projection image, the most similar matching objects are selected for each fluorescence signal region of the first fluorescence projection image; all matching objects are combined with their corresponding regions to form a preliminary pairing result set; in addition, if there are multiple regions that meet the conditions, the region with the highest overlap in spatial distribution features and lateral offset features is selected.

[0071] Furthermore, the initial pairing result set undergoes multiple constraint verification, sequentially executing intensity consistency constraints, geometric projection constraints, and feature consistency constraints, filtering out matching pairs that do not meet the conditions in stages, and gradually obtaining the fluorescence signal set.

[0072] In this embodiment of the invention, multiple constraint verifications are performed on each matching pair in the preliminary pairing result set to obtain a set of fluorescence signals that pass the verification. This includes: applying intensity consistency constraints to each matching pair in the preliminary pairing result set to obtain an initial set of fluorescence signals that pass the constraints; applying geometric projection constraints to each matching pair in the initial set of fluorescence signals to obtain an intermediate set of fluorescence signals that pass the constraints; and applying morphological feature consistency constraints to each matching pair in the intermediate set of fluorescence signals to obtain a set of fluorescence signals that pass the verification.

[0073] In this embodiment of the invention, during the process of applying intensity consistency constraints to each pair in the preliminary pairing result set, the signal intensity value of each pair in the preliminary pairing result set in their respective images is first obtained. The ratio or difference between these two intensity values ​​is calculated and compared with a preset reasonable fluctuation range. If the ratio or difference falls within the allowable tolerance range, the pairing point is considered to have passed the intensity consistency check. All pairs that pass this check are retained to form the initial fluorescence signal set.

[0074] Furthermore, in the process of geometrically projecting constraints on each matching pair in the initial fluorescence signal set, firstly, for each matching pair in the initial fluorescence signal set, the lateral position coordinates of the matching pair in the two images are extracted, and the actual displacement vector is calculated based on the lateral position coordinates in the two images. Then, the actual displacement vector is compared with the preset theoretical displacement vector. Only when the direction and magnitude of the actual displacement vector are within the error tolerance of the preset theoretical displacement vector is the matching pair determined to be constrained by geometric projection, and the constrained pairings constitute the intermediate fluorescence signal set.

[0075] Furthermore, in the process of constraining the morphological features of each matching pair in the intermediate fluorescence signal set, firstly, for each matching pair in the intermediate fluorescence signal set, the fine morphology of the corresponding region of the matching pair is extracted from the second fluorescence projection image. The fine morphology refers to the intensity distribution of the main lobe and the side lobes. The main lobe refers to the core region where the energy of the second non-diffraction beam is most concentrated, and the fluorescence signal intensity generated by the excited sample is the strongest. It appears as the brightest spot region on the second fluorescence projection image. The side lobes refer to the secondary regions surrounding the main lobe with lower energy than the main lobe. The fluorescence signal intensity generated by the excited sample is weaker than that of the main lobe. It appears as a spot or halo with a lower brightness than the main lobe on the image. Then, it is determined whether the side lobe is stably located directly above or below the main lobe based on the intensity distribution. Then, the observed actual side lobe orientation is compared with the preset theoretical orientation. If the two are consistent, the matching pair is determined to have passed the morphological feature consistency check; otherwise, it is excluded.

[0076] In this embodiment of the invention, spatial distribution features and lateral offset features are extracted without additional scanning and sampling, avoiding the efficiency bottleneck of traditional layer-by-layer scanning. This provides dual discrimination criteria for signal pairing, including axial and lateral features, thus improving the pairing discrimination. The spatial distribution feature vector and lateral offset vector are quantized into standardized vector form, eliminating the difference in the dimensions of the feature dimensions and ensuring the fairness and accuracy of feature comparison. By combining and generating regional feature descriptions, the limitations of single feature matching are avoided, significantly reducing the probability of mismatching of signals from the same source. By calculating similarity and matching to form a preliminary pairing result set, a preliminary pairing result set is formed, replacing traditional manual screening or low-dimensional matching methods, significantly improving pairing efficiency. At the same time, the optimal matching object is locked by similarity ranking, laying the foundation for accurate pairing. By performing multiple constraint verifications on the preliminary pairing result set, erroneous results in the preliminary pairing are filtered out, further improving the pairing accuracy and avoiding interference of invalid data with imaging quality.

[0077] S4. Stack the fluorescence signal sets into a three-dimensional fluorescence image corresponding to the sample according to the preset stacking rules.

[0078] In this embodiment of the invention, three-dimensional fluorescence images can achieve rapid, non-invasive volumetric observation and analysis of the internal three-dimensional structure of a sample without axial mechanical scanning.

[0079] In this embodiment of the invention, a set of fluorescence signals is stacked into a three-dimensional fluorescence image corresponding to the sample according to a preset stacking rule. This includes: performing coordinate inversion on each matching pair within the fluorescence signal set to obtain the three-dimensional spatial coordinates corresponding to each matching pair; selecting a fluorescence signal region belonging to the first fluorescence projection image from each matching pair within the fluorescence signal set, and determining the signal intensity value of the fluorescence region based on the calibration axial characteristics of the selected fluorescence region; combining the three-dimensional spatial coordinates and signal intensity values ​​into three-dimensional point cloud data according to the preset stacking rule; mapping each point cloud data to a corresponding voxel in a preset three-dimensional voxel grid to obtain a mapped voxel corresponding to each point cloud data; filling the mapped voxel with the signal intensity values ​​of the point cloud data to obtain a filled voxel grid; performing spatial interpolation calculation on empty voxels with empty intensity values ​​in the filled voxel grid to generate complete three-dimensional intensity volume data; and visually rendering the complete three-dimensional intensity volume data to obtain a three-dimensional fluorescence image corresponding to the sample.

[0080] In this embodiment of the invention, during the coordinate inversion process for each matching pair within the fluorescence signal set, each matching pair includes a fluorescence region of the first fluorescence projection image and a fluorescence region of the second fluorescence projection image. The abscissa of the fluorescence region belonging to the first fluorescence projection image and the abscissa of the fluorescence region belonging to the second fluorescence projection image in each matching pair within the fluorescence signal set are determined. The two abscissas are offset to obtain the calculated offset. The lateral displacement offset-axial position mapping table in step S2 is consulted to obtain the axial position corresponding to the calculated offset. The abscissa of the signal region in the first fluorescence projection image and the mapped axial position are combined to form a three-dimensional spatial coordinate.

[0081] Furthermore, the fluorescent region belonging to the first fluorescence projection image is first selected and associated with the calibration axis feature in step S1 to obtain the calibration axis feature of the fluorescent region belonging to the first fluorescence projection image. Step S1 has already stated that the calibration axis feature reflects the actual fluorescence signal intensity of the sample, so the signal intensity value of the fluorescent region belonging to the first fluorescence projection image can be determined based on the calibration axis feature.

[0082] Furthermore, the preset stacking rule is a standardized algorithm or spatial mapping criterion for systematically organizing, filling, and synthesizing the fluorescence signal set into continuous three-dimensional image data. The voxel grid is a discretized representation that divides three-dimensional space into regular small cubes (voxels). Through coordinate transformation algorithm, each point cloud data in the three-dimensional point cloud data is mapped to the corresponding voxel position, and the signal intensity value of the point cloud data is assigned to the voxel, ultimately forming a filled voxel grid.

[0083] Furthermore, during the spatial interpolation of empty voxels with empty intensity values ​​in the filled voxel grid, the intensity value of the empty voxels is estimated based on the intensity distribution pattern of the filled voxels around the empty voxels (such as the intensity gradient of neighboring voxels and the regional average intensity) by analyzing the intensity distribution pattern.

[0084] For example, if there are multiple high-intensity voxels around an empty voxel, the voxel is filled by weighted averaging of its neighborhood intensity values ​​to ensure that the intensity distribution of the entire voxel grid is continuous and conforms to the actual fluorescence characteristics of the sample, thus avoiding discontinuities or abnormal mutations.

[0085] Furthermore, by using volume rendering technology, the intensity values ​​of the complete three-dimensional intensity volume data are mapped to color and transparency to construct a three-dimensional scene with a sense of depth. For example, high-intensity areas can be rendered as light colors, and low-intensity areas as dark colors. Combined with a preset lighting model, the scattering and attenuation effects of fluorescence inside the sample are simulated, and finally a three-dimensional fluorescence image is generated, which facilitates intuitive observation and analysis of the three-dimensional fluorescence characteristics of the sample.

[0086] In this embodiment of the invention, a spatial framework for a three-dimensional image is established by mapping each point cloud data to a corresponding voxel in a preset three-dimensional voxel grid, thereby achieving an orderly distribution of signal intensity. By filling in the empty voxel intensity values, data gaps and missing values ​​are eliminated, ensuring the spatial continuity and integrity of the three-dimensional data. A three-dimensional fluorescence image is obtained through visualization rendering, transforming the abstract three-dimensional intensity volume data into an intuitive and analyzable three-dimensional image, thereby enabling precise observation of the fluorescence structure inside the sample.

[0087] In this embodiment of the invention, spatial interpolation calculation is performed on empty voxels with empty intensity values ​​in the filled voxel grid to generate complete three-dimensional intensity volume data. This includes: extracting filled voxels within a preset neighborhood from the filled voxel grid based on the empty voxels with empty intensity values; calculating the spatial distance between the empty voxels and the filled voxels, and determining the interpolation weight of the filled voxels based on the spatial distance; linearly calculating the interpolation weight and the intensity value of the filled voxels to obtain the interpolated intensity value of the empty voxels; and updating the filled voxel grid based on the interpolated intensity value to obtain complete three-dimensional intensity volume data.

[0088] In this embodiment of the invention, there may be one or more empty voxels with empty intensity values, and there may be one or more filled voxels. In the three-dimensional voxel grid, with the empty voxel with empty intensity values ​​as the center, all voxels within its preset neighborhood range are searched. Based on the spatial position relationship, it is determined which voxels are located in the neighboring region of the empty voxel (such as adjacent cubes or voxels within a specific radius), and voxels containing intensity values ​​are selected from the determined voxels, that is, filled voxels.

[0089] Furthermore, in the process of calculating the spatial distance between empty voxels and filled voxels, the coordinates of the empty voxel in its three-dimensional voxel grid and the coordinates of the filled voxels are calculated using the Euclidean distance formula to obtain the straight-line distance from the empty voxel to each of the surrounding filled voxels, and the straight-line distance is used as the spatial distance.

[0090] Furthermore, the interpolation weights of each filled voxel to the intensity value of the empty voxel are assigned according to the spatial distance; the closer the filled voxel is, the greater its influence on the empty voxel, and the higher its weight is assigned; the farther the filled voxel is, the smaller its influence, and the lower its weight is assigned accordingly.

[0091] Furthermore, the intensity values ​​of the filled voxels and the interpolation weights are linearly combined using a weighted average method. The result of the combination is used as the interpolated intensity value of the empty voxels, so that the interpolated intensity value can reflect the intensity distribution characteristics of the local area and maintain the smoothness of the spatial transition. Then, the interpolated intensity value is assigned to the corresponding empty voxel to complete the intensity value supplementation of all empty voxels in the filled voxel grid, and complete three-dimensional intensity volume data is obtained.

[0092] like Figure 4The diagram shown is a functional block diagram of a three-dimensional fluorescence imaging device based on a diffraction-free beam, provided in an embodiment of the present invention.

[0093] The three-dimensional fluorescence imaging device 400 based on a diffraction-free beam of the present invention can be installed in an electronic device. Depending on the functions implemented, the three-dimensional fluorescence imaging device 400 based on a diffraction-free beam may include an axial projection information extraction module 401, a lateral displacement information extraction module 402, a fluorescence information set pairing module 403, and a fluorescence image stacking module 404. The modules of the present invention can also be referred to as units, which refer to a series of computer program segments that can be executed by the processor of an electronic device and can perform a fixed function, and are stored in the memory of the electronic device.

[0094] In this embodiment, the functions of each module / unit are as follows:

[0095] The axial projection information extraction module 401 is used to acquire the first fluorescence projection image and the second fluorescence projection image of the preset sample, and extract the axial projection information of the sample based on the first fluorescence projection image.

[0096] The lateral displacement information extraction module 402 is used to extract the lateral displacement information of the sample based on the second fluorescence projection image.

[0097] The fluorescence information set pairing module 403 is used to pair the first fluorescence projection image with the second fluorescence projection image according to the axial projection information and the lateral displacement offset information to obtain a fluorescence signal set.

[0098] The fluorescence image stacking module 404 is used to stack the fluorescence signal set into a three-dimensional fluorescence image corresponding to the sample according to a preset stacking rule.

[0099] In one embodiment, the axial projection information extraction module 401 is specifically used to identify the fluorescence signal region corresponding to the sample in the first fluorescence projection image; perform grayscale distribution analysis on the fluorescence signal region, and extract the axial projection grayscale features of the fluorescence signal region based on the analysis results; calibrate the signal intensity of the axial projection grayscale features according to preset acquisition parameters to obtain calibrated axial features; and associate the calibrated axial features with the corresponding fluorescence signal region to generate axial projection information corresponding to the fluorescence signal region.

[0100] In one embodiment, the lateral displacement offset information extraction module 402 is specifically used to identify the homologous fluorescence signal region in the second fluorescence projection image corresponding to the fluorescence signal region; determine the second lateral coordinate of the fluorescence signal region in the second fluorescence projection image based on the homologous fluorescence signal region; extract the first lateral coordinate of the fluorescence signal region in the first fluorescence projection image; calculate the difference between the first lateral coordinate and the second lateral coordinate, and use the difference as the lateral displacement offset of the fluorescence signal region; and calculate the lateral displacement offset information corresponding to the fluorescence signal region using a preset displacement-axial position mapping formula based on a preset lateral coordinate constant and the lateral displacement offset.

[0101] In one embodiment, the fluorescence information set pairing module 403 is specifically used to extract the spatial distribution features of the fluorescence signal region from the axial projection information and extract the lateral displacement features of the fluorescence signal region from the lateral displacement information; convert the spatial distribution features and lateral displacement features into spatial distribution feature vectors corresponding to the distribution features and lateral displacement vectors corresponding to the lateral displacement features, respectively; determine the region feature description corresponding to the fluorescence signal region based on the spatial distribution feature vector, the lateral displacement vector, and the first lateral coordinate and the second lateral coordinate; calculate the feature similarity of the fluorescence signal region in the first fluorescence projection image and the second fluorescence projection image based on the region feature description; match the fluorescence signal region in the first fluorescence projection image with the fluorescence signal region in the second fluorescence projection image with the highest similarity based on the feature similarity to form a preliminary pairing result set; and perform multiple constraint verification on the preliminary pairing result set to obtain a fluorescence signal set that passes the verification.

[0102] In one embodiment, the fluorescence information set pairing module 403 is specifically used to apply intensity consistency constraints to each matching pair in the preliminary pairing result set to obtain an initial fluorescence signal set that passes the constraints; apply geometric projection constraints to each matching pair in the initial fluorescence signal set to obtain an intermediate fluorescence signal set that passes the constraints; and apply morphological feature consistency constraints to each matching pair in the intermediate fluorescence signal set to obtain a fluorescence signal set that passes the verification.

[0103] In one embodiment, the fluorescence image stacking module 404 is specifically used to perform coordinate inversion on each matching pair within the fluorescence signal set to obtain the three-dimensional spatial coordinates corresponding to each matching pair; select a fluorescence signal region belonging to the first fluorescence projection image from each matching pair within the fluorescence signal set, and determine the signal intensity value of the fluorescence region based on the calibration axial characteristics of the selected fluorescence region; combine the three-dimensional spatial coordinates and signal intensity values ​​into three-dimensional point cloud data according to a preset stacking rule; map each point cloud data in the three-dimensional point cloud data to the corresponding voxel in a preset three-dimensional voxel grid to obtain the mapped voxel corresponding to each point cloud data; fill the mapped voxel with the signal intensity value of the point cloud data to obtain a filled voxel grid; perform spatial interpolation calculation on the empty voxels with empty intensity values ​​in the filled voxel grid to generate complete three-dimensional intensity volume data; and perform visualization rendering on the complete three-dimensional intensity volume data to obtain the three-dimensional fluorescence image corresponding to the sample.

[0104] In one embodiment, the fluorescence image stacking module 404 is specifically used to extract filled voxels within a preset range from the filled voxel grid based on empty voxels with empty intensity values; calculate the spatial distance between empty voxels and filled voxels, and determine the interpolation weight of the filled voxels based on the spatial distance; perform linear calculation between the interpolation weight and the intensity value of the filled voxels to obtain the interpolated intensity value of the empty voxels; and update the filled voxel grid based on the interpolated intensity value to obtain complete three-dimensional intensity volume data.

[0105] In detail, each module in the three-dimensional fluorescence imaging device 400 based on a non-diffraction beam in this embodiment of the invention uses the same technical means as the three-dimensional fluorescence imaging method based on a non-diffraction beam in the accompanying drawings, and can produce the same technical effect, which will not be repeated here.

[0106] like Figure 5 The diagram shown is a schematic representation of an electronic device for implementing a three-dimensional fluorescence imaging method based on a diffraction-free beam, according to an embodiment of the present invention.

[0107] Electronic device 1 may include processor 10, memory 11, communication bus 12 and communication interface 13, and may also include computer programs stored in memory 11 and run on processor 10, such as a three-dimensional fluorescence imaging program based on a non-diffraction beam.

[0108] In some embodiments, the processor 10 may be composed of integrated circuits, such as a single packaged integrated circuit or multiple integrated circuits with the same or different functions, including combinations of one or more central processing units (CPUs), microprocessors, digital processing chips, graphics processors, and various control chips. The processor 10 is the control unit of the electronic device, connecting various components of the entire electronic device through various interfaces and lines. It executes programs or modules stored in the memory 11 (e.g., executing a three-dimensional fluorescence imaging program based on a non-diffractive beam) and calls data stored in the memory 11 to perform various functions of the electronic device and process data.

[0109] The memory 11 includes at least one type of readable storage medium, including flash memory, portable hard drive, multimedia card, card-type memory (e.g., SD or DX memory), magnetic memory, magnetic disk, optical disk, etc. In some embodiments, the memory 11 can be an internal storage unit of an electronic device, such as a portable hard drive. In other embodiments, the memory 11 can be an external storage device of the electronic device, such as a plug-in portable hard drive, smart media card (SMC), secure digital (SD) card, flash card, etc. Furthermore, the memory 11 can include both internal and external storage units of the electronic device. The memory 11 can be used not only to store application software and various types of data installed on the electronic device, such as code for a three-dimensional fluorescence imaging program based on a diffraction-free beam, but also to temporarily store data that has been output or will be output.

[0110] The communication bus 12 can be a Peripheral Component Interconnect (PCI) bus or an Extended Industry Standard Architecture (EISA) bus, etc. This bus can be divided into an address bus, a data bus, a control bus, etc. The bus is configured to enable communication between the memory 11 and at least one processor 10, etc.

[0111] Communication interface 13 is used for communication between the aforementioned electronic device and other devices, including a network interface and a user interface. Optionally, the network interface may include a wired interface and / or a wireless interface (such as a Wi-Fi interface, Bluetooth interface, etc.), typically used to establish communication connections between the electronic device and other electronic devices. The user interface may be a display, an input unit (such as a keyboard), and optionally, a standard wired or wireless interface. Optionally, in some embodiments, the display may be an LED display, a liquid crystal display, a touch-sensitive liquid crystal display, or an OLED (Organic Light-Emitting Diode) touchscreen, etc. The display may also be appropriately referred to as a screen or display unit, used to display information processed in the electronic device and to display a visual user interface.

[0112] Figure 5 Only electronic devices with components are shown; it will be understood by those skilled in the art that... Figure 5 The structure shown does not constitute a limitation on the electronic device 1, and may include fewer or more components than shown, or combine certain components, or have different component arrangements.

[0113] For example, although not shown, the electronic device may also include a power supply (such as a battery) to power various components. Preferably, the power supply can be logically connected to at least one processor 10 via a power management device, thereby enabling functions such as charging management, discharging management, and power consumption management. The power supply may also include one or more DC or AC power sources, recharging devices, power fault detection circuits, power converters or inverters, power status indicators, and other arbitrary components. The electronic device may also include various sensors, Bluetooth modules, Wi-Fi modules, etc., which will not be described in detail here.

[0114] It should be understood that the embodiments are for illustrative purposes only and are not limited to this structure in the scope of the patent application.

[0115] Specifically, the processor 10's specific implementation method of the above instructions can be found in the description of the relevant steps in the corresponding embodiments of the accompanying drawings, and will not be repeated here.

[0116] Furthermore, if the modules / units integrated in electronic device 1 are implemented as software functional units and sold or used as independent products, they can be stored in a computer-readable storage medium. The computer-readable storage medium can be volatile or non-volatile. For example, a computer-readable medium may include: any entity or device capable of carrying computer program code, a recording medium, a USB flash drive, a portable hard drive, a magnetic disk, an optical disk, a computer memory, or a read-only memory (ROM).

[0117] The present invention also provides a computer-readable storage medium storing a computer program. When executed by a processor, the computer program can implement a three-dimensional fluorescence imaging method based on a diffraction-free beam according to any of the above embodiments. It should be noted that the computer-readable storage medium can be volatile or non-volatile. For example, a computer-readable medium may include: any entity or device capable of carrying computer program code, a recording medium, a USB flash drive, a portable hard drive, a magnetic disk, an optical disk, a computer memory, or a read-only memory (ROM).

[0118] In the several embodiments provided by this invention, it should be understood that the disclosed devices, apparatuses, and methods can be implemented in other ways. For example, the apparatus embodiments described above are merely illustrative; for instance, the division of modules is only a logical functional division, and other division methods may be used in actual implementation.

[0119] The modules described as separate components may or may not be physically separate. The components shown as modules may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the modules can be selected to achieve the purpose of this embodiment according to actual needs.

[0120] Furthermore, the functional modules in the various embodiments of the present invention can be integrated into one processing unit, or each unit can exist physically separately, or two or more units can be integrated into one unit. The integrated unit can be implemented in hardware or in the form of hardware plus software functional modules.

[0121] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention.

[0122] Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be embraced within the invention. No appended diagram markings in the claims should be construed as limiting the scope of the claims.

[0123] Furthermore, it is clear that the word "comprising" does not exclude other units or steps, and the singular does not exclude the plural. Multiple units or devices recited in a system claim may also be implemented by a single unit or device through software or hardware. The terms "first," "second," etc., are used to indicate names and do not indicate any specific order.

[0124] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A three-dimensional fluorescence imaging method based on a diffraction-free beam, characterized in that, The method includes: Acquire a first fluorescence projection image and a second fluorescence projection image of a preset sample, and extract the axial projection information of the sample based on the first fluorescence projection image; The lateral displacement information of the sample is extracted based on the second fluorescence projection image; Based on the axial projection information and the lateral displacement offset information, the first fluorescence projection image and the second fluorescence projection image are paired to obtain a set of fluorescence signals; The fluorescence signal set is stacked into a three-dimensional fluorescence image corresponding to the sample according to a preset stacking rule; The step of extracting the axial projection information of the sample based on the first fluorescence projection image includes: Identify the fluorescence signal region in the first fluorescence projection image that corresponds to the sample; Gray-scale distribution analysis is performed on the fluorescence signal region, and the axial projection gray-scale features of the fluorescence signal region are extracted based on the analysis results; The axial projection grayscale features are calibrated by signal intensity according to preset acquisition parameters to obtain calibrated axial features; The calibration axial feature is associated with the corresponding fluorescence signal region to generate axial projection information corresponding to the fluorescence signal region; The step of extracting the lateral displacement information of the sample based on the second fluorescence projection image includes: Identify the homologous fluorescence signal region in the second fluorescence projection image that corresponds to the fluorescence signal region; The second lateral coordinate of the fluorescence signal region in the second fluorescence projection image is determined based on the homologous fluorescence signal region. Extract the first lateral coordinate of the fluorescence signal region in the first fluorescence projection image; Calculate the difference between the first horizontal coordinate and the second horizontal coordinate, and use the difference as the horizontal displacement offset of the fluorescence signal region; The lateral displacement offset information corresponding to the fluorescence signal region is obtained by using a preset displacement-axial position mapping formula based on a preset horizontal coordinate constant and the lateral displacement offset. The step of pairing the first fluorescence projection image with the second fluorescence projection image based on the axial projection information and the lateral displacement offset information to obtain a fluorescence signal set includes: The spatial distribution features of the fluorescence signal region are extracted from the axial projection information, and the lateral displacement features of the fluorescence signal region are extracted from the lateral displacement information. The spatial distribution feature and the lateral offset feature are respectively converted into a spatial distribution feature vector corresponding to the distribution feature and a lateral offset vector corresponding to the lateral offset feature; The region feature description corresponding to the fluorescence signal region is determined based on the spatial distribution feature vector, the lateral offset vector, the first lateral coordinate, and the second lateral coordinate. Based on the regional feature description, the feature similarity of the fluorescence signal regions in the first fluorescence projection image and the second fluorescence projection image is calculated. Based on the feature similarity, the fluorescence signal region in the first fluorescence projection image is matched with the fluorescence signal region with the highest similarity in the second fluorescence projection image to form a preliminary pairing result set; The preliminary pairing result set is subjected to multiple constraint verifications to obtain a set of fluorescence signals that pass the verifications; The step of performing multiple constraint checks on the preliminary pairing result set to obtain a set of fluorescence signals that pass the checks includes: Each matching pair in the preliminary pairing result set is subjected to intensity consistency constraints to obtain an initial fluorescence signal set that passes the constraints; Geometric projection constraints are applied to each matching pair in the initial fluorescence signal set to obtain an intermediate fluorescence signal set that passes the constraints; A morphological feature consistency constraint is applied to each matching pair in the intermediate fluorescence signal set to obtain a verified fluorescence signal set.

2. The three-dimensional fluorescence imaging method based on a diffraction-free beam as described in claim 1, characterized in that, The step of stacking the fluorescence signal set into a three-dimensional fluorescence image corresponding to the sample according to a preset stacking rule includes: For each matching pair in the fluorescence signal set, coordinate inversion is performed to obtain the three-dimensional spatial coordinates corresponding to each matching pair; A fluorescence signal region belonging to the first fluorescence projection image is selected from each matching pair within the fluorescence signal set, and the signal intensity value of the selected fluorescence region is determined based on the calibration axial characteristics of the selected fluorescence region. The three-dimensional spatial coordinates and signal strength values ​​are combined into three-dimensional point cloud data according to the preset stacking rules; Each point cloud data in the three-dimensional point cloud data is mapped to a corresponding voxel in a preset three-dimensional voxel grid to obtain the mapped voxel corresponding to each point cloud data. The signal strength values ​​of the point cloud data are filled into the mapped voxels to obtain a filled voxel grid. Spatial interpolation calculations are performed on empty voxels with empty intensity values ​​in the filled voxel mesh to generate complete three-dimensional intensity volume data; The complete three-dimensional intensity volume data is visualized and rendered to obtain the three-dimensional fluorescence image corresponding to the sample.

3. The three-dimensional fluorescence imaging method based on a diffraction-free beam as described in claim 2, characterized in that, The step of performing spatial interpolation calculations on empty voxels with empty intensity values ​​in the filled voxel mesh to generate complete three-dimensional intensity volume data includes: Based on empty voxels with empty intensity values, fill voxels within a predetermined range are extracted from the filled voxel grid. Calculate the spatial distance between the empty voxel and the filled voxel, and determine the interpolation weight of the filled voxel based on the spatial distance; The interpolation weights are linearly calculated with the intensity values ​​of the filled voxels to obtain the interpolated intensity values ​​of the empty voxels. The infill voxel mesh is updated based on the interpolated intensity value to obtain complete three-dimensional intensity volume data.

4. A three-dimensional fluorescence imaging device based on a diffraction-free beam, characterized in that, The device includes: An axial projection information extraction module is used to acquire a first fluorescence projection image and a second fluorescence projection image of a preset sample, and extract the axial projection information of the sample based on the first fluorescence projection image. A lateral displacement information extraction module is used to extract the lateral displacement information of the sample based on the second fluorescence projection image; A fluorescence information set pairing module is used to pair the first fluorescence projection image with the second fluorescence projection image according to the axial projection information and the lateral displacement offset information to obtain a fluorescence signal set. A fluorescence image stacking module is used to stack the fluorescence signal set into a three-dimensional fluorescence image corresponding to the sample according to a preset stacking rule; The axial projection information extraction module is specifically used for: identifying the fluorescence signal region in the first fluorescence projection image corresponding to the sample; performing grayscale distribution analysis on the fluorescence signal region, and extracting the axial projection grayscale features of the fluorescence signal region based on the analysis results; calibrating the signal intensity of the axial projection grayscale features according to preset acquisition parameters to obtain calibrated axial features; associating the calibrated axial features with the corresponding fluorescence signal region to generate axial projection information corresponding to the fluorescence signal region. The lateral displacement information extraction module is specifically used for: identifying a homologous fluorescence signal region in the second fluorescence projection image corresponding to the fluorescence signal region; determining the second lateral coordinate of the fluorescence signal region in the second fluorescence projection image based on the homologous fluorescence signal region; extracting the first lateral coordinate of the fluorescence signal region in the first fluorescence projection image; calculating the difference between the first lateral coordinate and the second lateral coordinate, and using the difference as the lateral displacement of the fluorescence signal region; and calculating the lateral displacement information corresponding to the fluorescence signal region using a preset displacement-axial position mapping formula based on a preset lateral coordinate constant and the lateral displacement. The fluorescence information set pairing module is specifically used for: extracting the spatial distribution features of the fluorescence signal region from the axial projection information, and extracting the lateral displacement features of the fluorescence signal region from the lateral displacement information; converting the spatial distribution features and the lateral displacement features into spatial distribution feature vectors and lateral displacement vectors corresponding to the distribution features and the lateral displacement features, respectively; determining the region feature description corresponding to the fluorescence signal region based on the spatial distribution feature vector, the lateral displacement vector, the first lateral coordinate, and the second lateral coordinate; calculating the feature similarity of the fluorescence signal regions in the first fluorescence projection image and the second fluorescence projection image based on the region feature description; matching the fluorescence signal region in the first fluorescence projection image with the fluorescence signal region in the second fluorescence projection image with the highest similarity based on the feature similarity to form a preliminary pairing result set; applying intensity consistency constraints to each matching pair in the preliminary pairing result set to obtain an initial fluorescence signal set that passes the constraints; applying geometric projection constraints to each matching pair in the initial fluorescence signal set to obtain an intermediate fluorescence signal set that passes the constraints; and applying morphological feature consistency constraints to each matching pair in the intermediate fluorescence signal set to obtain a verified fluorescence signal set.

5. An electronic device, characterized in that, The electronic device includes: At least one processor; and, A memory communicatively connected to the at least one processor; wherein, The memory stores a computer program that can be executed by the at least one processor, the computer program being executed by the at least one processor to enable the at least one processor to perform the three-dimensional fluorescence imaging method based on a diffraction-free beam as described in any one of claims 1 to 3.

6. A computer-readable storage medium storing a computer program, characterized in that, When the computer program is executed by the processor, it implements the three-dimensional fluorescence imaging method based on a non-diffraction beam as described in any one of claims 1 to 3.

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