Digital adaptive optical method and apparatus for multiphoton imaging

Abstract

The application relates to the technical field of microscopic imaging, in particular to a digital adaptive optical method and device for multi-photon imaging, wherein the method comprises the following steps: based on a two-photon synthetic aperture microscope, three-dimensional sample projections are collected by using a small-aperture excitation light beam, small-aperture imaging results of different viewing angles are generated, and a lateral shift vector of images of different sub-apertures caused by wavefront distortion phase in an imaging process is calculated, and in the process of obtaining a three-dimensional reconstruction result based on three-dimensional sample information, optical distortion is compensated according to the lateral shift vector. According to the embodiment of the application, the lateral shift vector generated by each small-aperture image is used to compensate for system optical distortion and perform three-dimensional reconstruction of sample information, so that multi-photon imaging with high resolution and high volume imaging speed is realized, the robustness and practicability of the optical system are improved, and the imaging efficiency is higher.

Description

Technical Field

[0001] This application relates to the field of microscopic imaging technology, and in particular to a digital adaptive optics method and apparatus for multiphoton imaging. Background Technology

[0002] With the continuous development of laser technology, multiphoton microscopy has been widely used in more and more life science research. Multiphoton microscopy has a deep penetration depth and strong tomographic ability, and can be used for imaging deep scattering samples.

[0003] In related technologies, the image quality of multiphoton imaging microscopes will be significantly reduced in the presence of optical distortion in the optical system or tissue scattering in three-dimensional samples. Adaptive optics can compensate for system distortion by measuring the magnitude of optical system distortion during imaging.

[0004] However, in related technologies, the imaging environment needs to be measured in advance before the distorted phase is obtained, which leads to a decrease in the imaging speed of the system. Furthermore, the process of compensating for wavefront distortion relies on active optical devices, which increases the complexity of the optical system and makes it impossible to quickly achieve high-quality multiphoton imaging. This reduces the imaging efficiency of the optical system and its practicality is low, which urgently needs to be addressed. Summary of the Invention

[0005] This application provides a digital adaptive optics method and apparatus for multiphoton imaging to solve the problems in related technologies, such as the need to measure the imaging environment in advance before acquiring the distorted phase, which leads to a decrease in the system imaging speed, and the reliance on active optical devices for wavefront distortion compensation, which increases the complexity of the optical system, makes it impossible to quickly achieve high-quality multiphoton imaging, reduces the imaging efficiency of the optical system, and results in low practicality.

[0006] The first aspect of this application provides a digital adaptive optics method for multiphoton imaging, comprising the following steps: based on a two-photon synthetic aperture microscope, using a small-aperture excitation beam constrained by a preset diffraction-limited aperture, acquiring three-dimensional sample projections from different viewpoints to generate small-aperture imaging results from different viewpoints; based on the small-aperture imaging results, calculating the lateral offset vectors generated by wavefront distortion phase caused by system distortion or tissue scattering during the imaging process, resulting in images with different sub-apertures, and obtaining three-dimensional sample information that meets preset resolution conditions; and in the process of obtaining three-dimensional reconstruction results based on the three-dimensional sample information, compensating for optical distortion according to the lateral offset vector.

[0007] Furthermore, in one embodiment of this application, the process of obtaining the three-dimensional reconstruction result based on the three-dimensional sample information, and compensating for optical distortion according to the lateral offset vector, includes: applying the wavefront distortion phase determined by the lateral offset vector to the generation of the system point spread function in the preset three-dimensional reconstruction algorithm.

[0008] Specifically, in one embodiment of this application, the lateral offset vector and the derivative function of the wavefront distortion phase satisfy the following relationship:

[0009]

[0010] Where Δφ is the amount of distortion phase change, and φ(x,y) is the distorted incident light wavefront function. The offset vector on the X-axis of the image for each small aperture image. This is the offset vector on the Y-axis of the image for each small aperture image.

[0011] Furthermore, in one embodiment of this application, the process of obtaining the three-dimensional reconstruction result based on the three-dimensional sample information, and compensating for optical distortion according to the lateral offset vector, includes: compensating for the lateral offset vector in the preset three-dimensional reconstruction algorithm, and then performing three-dimensional reconstruction.

[0012] A second aspect of this application provides a digital adaptive optics device for multiphoton imaging, comprising: an acquisition module, configured to acquire three-dimensional sample projections from different perspectives using a small-aperture excitation beam constrained by a preset diffraction-limited aperture microscope based on a two-photon synthetic aperture microscope, thereby generating small-aperture imaging results from different perspectives; a calculation module, configured to calculate, based on the small-aperture imaging results, the lateral offset vectors generated by wavefront distortion phase caused by system distortion or tissue scattering during the imaging process, resulting in images with different sub-apertures, and to obtain three-dimensional sample information that meets preset resolution conditions; and an imaging module, configured to compensate for optical distortion based on the lateral offset vector during the process of obtaining three-dimensional reconstruction results based on the three-dimensional sample information.

[0013] Furthermore, in one embodiment of this application, the imaging module includes: a correction unit, used to apply the wavefront distortion phase determined by the lateral offset vector to the generation of the system point spread function in a preset three-dimensional reconstruction algorithm.

[0014] Specifically, in one embodiment of this application, the lateral offset vector and the derivative function of the wavefront distortion phase satisfy the following relationship:

[0015]

[0016] Where Δφ is the amount of distortion phase change, and φ(x,y) is the distorted incident light wavefront function. The offset vector on the X-axis of the image for each small aperture image. This is the offset vector on the Y-axis of the image for each small aperture image.

[0017] Furthermore, in one embodiment of this application, the imaging module includes a compensation unit, used to compensate for the lateral offset vector in the preset three-dimensional reconstruction algorithm before performing three-dimensional reconstruction.

[0018] A third aspect of this application provides an electronic device, including: a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the processor executes the program to implement the digital adaptive optics method for multiphoton imaging as described in the above embodiments.

[0019] A fourth aspect of this application provides a computer-readable storage medium storing a computer program that, when executed by a processor, implements the digital adaptive optics method for multiphoton imaging as described above.

[0020] This application's embodiments utilize digital adaptive optics to compensate for system optical distortion and perform three-dimensional reconstruction of sample information based on the lateral offset vectors generated from each small-aperture image. This achieves high-resolution and high-volume imaging speed multiphoton imaging, improving the robustness and practicality of the optical system and increasing imaging efficiency. Therefore, it solves the problems in related technologies, such as the need to measure the imaging environment before acquiring the distorted phase, leading to a decrease in system imaging speed, and the reliance on active optical devices for wavefront distortion compensation, which increases the complexity of the optical system, hinders the rapid achievement of high-quality multiphoton imaging, reduces the imaging efficiency of the optical system, and results in low practicality.

[0021] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0022] The above and / or additional aspects and advantages of this application will become apparent and readily understood from the following description of the embodiments taken in conjunction with the accompanying drawings, wherein:

[0023] Figure 1 This is a flowchart of a digital adaptive optics method for multiphoton imaging provided according to an embodiment of this application;

[0024] Figure 2 This is a schematic diagram illustrating the process of digital adaptive optics calculating image offset caused by distortion according to an embodiment of this application;

[0025] Figure 3 This is a schematic diagram illustrating the usage principle of digital adaptive optics according to one embodiment of this application;

[0026] Figure 4 This image shows a result of multiphoton microscopy imaging of fluorescent microspheres using a digital adaptive optics method according to an embodiment of this application.

[0027] Figure 5 This image shows a mouse brain slice captured by multiphoton microscopy using a digital adaptive optics method, according to an embodiment of this application.

[0028] Figure 6 This is a schematic diagram illustrating the process of obtaining the distorted phase from the offset in a two-photon synthetic aperture microscope according to an embodiment of this application.

[0029] Figure 7 This is a schematic diagram illustrating the principle of wavefront distortion causing lateral shift in a sub-aperture image according to an embodiment of this application;

[0030] Figure 8 This is a schematic diagram of a digital adaptive optics device for multiphoton imaging according to an embodiment of this application;

[0031] Figure 9 This is a schematic diagram of the structure of an electronic device according to an embodiment of this application. Detailed Implementation

[0032] The embodiments of this application are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain this application, and should not be construed as limiting this application.

[0033] The following description, with reference to the accompanying drawings, describes a digital adaptive optics method and apparatus for multiphoton imaging according to embodiments of this application. Addressing the issues raised in the background art, where prior measurement of the imaging environment is required before acquiring the distorted phase, leading to a decrease in system imaging speed, and the wavefront distortion compensation process relies on active optical devices, increasing the complexity of the optical system and hindering the rapid achievement of high-quality multiphoton imaging, thus reducing the imaging efficiency and practicality of the optical system, this application provides a digital adaptive optics method for multiphoton imaging. Based on the lateral offset vector generated by each small-aperture image, the method compensates for system optical distortion and performs three-dimensional reconstruction of sample information, thereby achieving high-resolution and high-volume imaging speed multiphoton imaging, improving the robustness and practicality of the optical system, and increasing imaging efficiency. This solves the problems in the related art, such as the need for prior measurement of the imaging environment before acquiring the distorted phase, resulting in a decrease in system imaging speed, and the reliance on active optical devices for wavefront distortion compensation, which increases the complexity of the optical system, hinders the rapid achievement of high-quality multiphoton imaging, reduces the imaging efficiency, and reduces practicality.

[0034] Specifically, Figure 1This is a schematic flowchart of a digital adaptive optics method for multiphoton imaging provided in an embodiment of this application.

[0035] like Figure 1 As shown, the digital adaptive optics method for multiphoton imaging includes the following steps:

[0036] In step S101, based on the two-photon synthetic aperture microscope, a small aperture excitation beam constrained by a preset diffraction-limited aperture is used to collect three-dimensional sample projections from different perspectives, generating small aperture imaging results from different perspectives.

[0037] It is understood that the two-photon synthetic aperture microscope in this application embodiment can achieve image of the sample by synthesizing a large aperture through the process of sub-apertures. Multiple small apertures can be used to collect three-dimensional sample projections from different perspectives by the excitation beam. The image generated by each small aperture is obtained by the point spread function during system imaging, and small aperture imaging results from different perspectives are obtained.

[0038] It should be noted that the preset diffraction limit size is set by those skilled in the art based on the actual situation, and no specific limitation is made here.

[0039] The embodiments of this application can be based on a two-photon synthetic aperture microscope, using a small aperture excitation beam constrained by a preset diffraction-limited small aperture to acquire three-dimensional sample projections from different perspectives, generating small aperture imaging results from different perspectives, thereby providing relevant imaging information for further applications of digital adaptive optics in the following steps, and satisfying the data foundation required for distortion correction in the multiphoton imaging process.

[0040] In step S102, based on the small aperture imaging results, the lateral offset vectors generated by wavefront distortion phase caused by system distortion or tissue scattering during the imaging process are calculated, and three-dimensional sample information that meets the preset resolution conditions is obtained.

[0041] It is understood that optical system distortion or tissue scattering in the embodiments of this application can cause deformation of the wavefront of the incident excitation light, and the deformation of the incident light wavefront can cause lateral shift in images captured by small apertures at different positions. The small aperture imaging results obtained from the above steps can be processed by a three-dimensional reconstruction algorithm deployed on a two-photon synthesized aperture microscope to obtain three-dimensional sample information that meets the preset resolution conditions.

[0042] It should be noted that the preset resolution conditions are set by those skilled in the art according to the actual situation, and are not specifically limited here.

[0043] Specifically, the process of obtaining the lateral offset vector generated by different sub-aperture images is as follows: Figure 2 As shown. Figure 2(A) The obtained 3D data can be convolved with the point spread function of the image using a 3D reconstruction algorithm to obtain the forward projection P(x,y) of the original measurement viewpoint. A correlation matrix is ​​then calculated by correlating this forward projection with the original measurement data M(x,y). The maximum value of the correlation matrix represents the coordinates of the position with the highest correlation. The distance from the center position coordinates is (Shiftx, Shifty), from which the offset vector is obtained. and exist Figure 2 In (B), when the number of small apertures is 13, the calculation results of the offset distance of the three perspectives and the corresponding position and correlation curves are shown in the figure.

[0044] This application embodiment can calculate the lateral offset vector of images with different sub-apertures caused by wavefront distortion phase due to system distortion or tissue scattering during the imaging process based on the small aperture imaging results, and obtain three-dimensional sample information that meets the preset resolution conditions. This realizes the data processing process of digital adaptive optics method in two-photon synthetic aperture microscope application and improves the feasibility of multiphoton imaging distortion correction.

[0045] In step S103, during the process of obtaining the three-dimensional reconstruction result based on the three-dimensional sample information, optical distortion is compensated according to the lateral offset vector.

[0046] It is understood that the specific implementation process of compensating for optical distortion based on the lateral offset vector in the embodiments of this application does not require the shooting time during imaging. Distortion compensation is performed during the post-processing of imaging data, thereby obtaining a three-dimensional reconstruction result based on three-dimensional sample information and obtaining the final imaging image.

[0047] For example, such as Figure 3 The diagram illustrates the principle of digital adaptive optics according to an embodiment of this application. In traditional two-photon microscopy (TPM), image quality degrades when optical distortion or sample scattering occurs. However, in two-photon synthetic aperture microscopy (2pSAM), when detecting small-aperture imaging results from different perspectives, the image itself encodes the wavefront changes as it passes through optical distortion and scattering tissue during the small-aperture imaging process. Therefore, high-quality, high-resolution imaging images can be recovered through three-dimensional reconstruction by calculating and applying the digital adaptive optics (DAO) method.

[0048] like Figure 4The figures shown are images of fluorescent microspheres captured by a multiphoton microscope using a digital adaptive optics method according to an embodiment of this application. The digital adaptive optics method was implemented in the imaging experiment of the fluorescent microsphere sample. The right figure shows the normal imaging result obtained by a conventional two-photon microscope capturing a 0.2 μm diameter fluorescent microsphere. The left figure shows the imaging result of the two-photon microscope after introducing a mismatch correction loop, increasing the optical system distortion. The middle figure shows the final imaging result obtained by a two-photon synthetic aperture microscope combining the digital adaptive optics method, clearly demonstrating the improved distortion resistance of the multiphoton microscope.

[0049] like Figure 5 The image shown is a result of multiphoton microscopy imaging of mouse brain slices using a digital adaptive optics method according to an embodiment of this application. In the imaging experiment of mouse brain slice samples, the digital adaptive optics method was implemented. For imaging mouse brain slice samples with neurons expressing green fluorescent protein, two complex multiphoton imaging conditions were introduced: mismatched correction loops and multiphoton imaging under a water microscope objective without added wetting water. This was used to measure the effect of the digital adaptive optics method on improving the microscope's anti-distortion capability. The figure compares the imaging results of ordinary two-photon microscopy (TPM, w / o deconv), ordinary two-photon microscopy imaging results with full aperture point spread function deconvolution (TPM, w / deconv), two-photon synthetic aperture microscopy without using the digital adaptive optics method (2pSAM, w / o DAO), and two-photon synthetic aperture microscopy with the digital adaptive optics method (2pSAM, w / DAO). Figure 5 (E) is the image result taken by two-photon microscope under normal conditions, which serves as a reference for the true value. Figure 5 (B, D, F) are respectively Figure 5 The magnified view of (A, C, E) shows that the image quality of the two-photon aperture synthesis microscope combined with the digital adaptive optics method has been improved and restored, and the resolution has been recovered.

[0050] In the process of obtaining three-dimensional reconstruction results based on three-dimensional sample information, the embodiments of this application can compensate for optical distortion according to the lateral offset vector. By using digital adaptive optics in a two-photon synthetic aperture microscope, the imaging speed and imaging quality of the optical system are improved, thereby obtaining high-resolution multiphoton imaging results, which are more practical.

[0051] Furthermore, in one embodiment of this application, in the process of obtaining the three-dimensional reconstruction result based on the three-dimensional sample information, optical distortion is compensated according to the lateral offset vector, including: applying the wavefront distortion phase determined by the lateral offset vector to the generation of the system point spread function in the preset three-dimensional reconstruction algorithm.

[0052] It is understood that, in the embodiments of this application, the wavefront distortion phase can be calculated based on the relationship between the lateral offset vector and the derivative function corresponding to the wavefront distortion phase, thereby regenerating the corrected point spread function for system imaging. The corrected point spread function approximates the most realistic excitation point spread function during imaging. Then, through a three-dimensional reconstruction algorithm, the three-dimensional information of the sample is solved using the projections from different viewpoints obtained from the original measurement data and the phase-corrected point spread function, and the final sample imaging result is obtained.

[0053] It should be noted that the preset 3D reconstruction algorithm is set by those skilled in the art according to the actual situation, and no specific limitation is made here.

[0054] like Figure 6 The diagram shown is a schematic of the process of obtaining the aberration phase from the offset in a two-photon synthetic aperture microscope according to an embodiment of this application. The number of sub-apertures used in the two-photon synthetic aperture microscope is set to 13. The offset of the 13 sub-aperture images can be used to obtain the shift map in the X and Y directions of the large aperture formed by the sub-apertures. The specific distribution and size of the aberration phase of the corresponding large aperture can be calculated by integrating the shift map.

[0055] The embodiments of this application can apply the wavefront distortion phase determined by the lateral offset vector to the generation of the system point spread function in the preset three-dimensional reconstruction algorithm, thereby correcting the image quality degradation caused by optical distortion and improving the resolution of the final imaging result.

[0056] Specifically, in one embodiment of this application, the derivative function of the lateral offset vector and the wavefront distortion phase satisfies the following relationship:

[0057]

[0058] Where Δφ is the amount of distortion phase change, and φ(x,y) is the distorted incident light wavefront function. The offset vector on the X-axis of the image for each small aperture image. This is the offset vector on the Y-axis of the image for each small aperture image.

[0059] Understandably, according to wave optics theory, the translation of image information below the objective lens is the derivative function of the wavefront distortion of the objective lens aperture plane. Therefore, the phase magnitude of the wavefront distortion of the incident light can be obtained by integrating the lateral offset of the small aperture imaging result.

[0060] like Figure 7This is a schematic diagram illustrating the principle of wavefront distortion causing lateral shift in sub-aperture images according to an embodiment of this application. The incident light wavefront function is φ0(x,y), which becomes φ(x,y) after optical distortion of the optical system or tissue scattering of the three-dimensional sample. Each small aperture corresponds to a different lateral shift, and the relationship between the lateral shift vector and the derivative function corresponding to the wavefront distortion phase is shown in the formula in the figure.

[0061] Furthermore, in one embodiment of this application, in the process of obtaining the three-dimensional reconstruction result based on the three-dimensional sample information, optical distortion is compensated according to the lateral offset vector, including: compensating the lateral offset vector in a preset three-dimensional reconstruction algorithm, and then performing three-dimensional reconstruction.

[0062] It is understood that, in the embodiments of this application, the image offset vector obtained in the above steps can be used. and In the 3D reconstruction algorithm, the original measurement data is first compensated for the lateral offset. Then, the subsequent 3D reconstruction algorithm is carried out based on the compensated data to solve the 3D information of the sample. The offset vector is then applied to the next iteration of the 3D reconstruction algorithm.

[0063] The embodiments of this application can compensate for the lateral offset vector in the preset three-dimensional reconstruction algorithm before performing three-dimensional reconstruction, thereby improving the resolution and quality of the image and making multiphoton imaging more practical.

[0064] The digital adaptive optics method for multiphoton imaging proposed in this application can compensate for system optical distortion and perform three-dimensional reconstruction of sample information based on the lateral offset vector generated by each small-aperture image, thereby achieving high-resolution and high-volume imaging speed multiphoton imaging. This improves the robustness and practicality of the optical system and increases imaging efficiency. Therefore, it solves the problems in related technologies, such as the need to measure the imaging environment before acquiring the distorted phase, which leads to a decrease in system imaging speed, and the reliance on active optical devices in the wavefront distortion compensation process, which increases the complexity of the optical system, makes it impossible to quickly achieve high-quality multiphoton imaging, reduces the imaging efficiency of the optical system, and results in low practicality.

[0065] Next, with reference to the accompanying drawings, a digital adaptive optics device for multiphoton imaging according to an embodiment of this application is described.

[0066] Figure 8 This is a schematic diagram of the structure of a digital adaptive optics device for multiphoton imaging according to an embodiment of this application.

[0067] like Figure 8 As shown, the digital adaptive optics device 10 for multiphoton imaging includes: an acquisition module 100, a computing module 200, and an imaging module 300.

[0068] The acquisition module 100 is used to acquire three-dimensional sample projections from different perspectives and generate small-aperture imaging results from different perspectives using a small-aperture excitation beam constrained by a pre-set diffraction-limited aperture microscope based on a two-photon synthetic aperture microscope.

[0069] The calculation module 200 is used to calculate the lateral offset vector of the image with different sub-apertures caused by wavefront distortion phase caused by system distortion or tissue scattering during the imaging process based on the small aperture imaging results, and to obtain three-dimensional sample information that meets the preset resolution conditions.

[0070] The imaging module 300 is used to compensate for optical distortion based on the lateral offset vector during the process of obtaining the three-dimensional reconstruction result based on the three-dimensional sample information.

[0071] Furthermore, in one embodiment of this application, the imaging module 300 includes a correction unit.

[0072] The correction unit is used to apply the wavefront distortion phase determined by the lateral offset vector to the generation of the system point spread function in the preset three-dimensional reconstruction algorithm.

[0073] Specifically, in one embodiment of this application, the derivative function of the lateral offset vector and the wavefront distortion phase satisfies the following relationship:

[0074]

[0075] Where Δφ is the amount of distortion phase change, and φ(x,y) is the distorted incident light wavefront function. The offset vector on the X-axis of the image for each small aperture image. This is the offset vector on the Y-axis of the image for each small aperture image.

[0076] Furthermore, in one embodiment of this application, the imaging module 300 includes a compensation unit.

[0077] The compensation unit is used to compensate for the lateral offset vector in the preset 3D reconstruction algorithm before performing 3D reconstruction.

[0078] It should be noted that the foregoing explanation of the digital adaptive optics method embodiment for multiphoton imaging also applies to the digital adaptive optics device for multiphoton imaging in this embodiment, and will not be repeated here.

[0079] The digital adaptive optics device for multiphoton imaging proposed in this application can compensate for system optical distortion and perform three-dimensional reconstruction of sample information based on the lateral offset vector generated by each small-aperture image, thereby achieving high-resolution and high-volume imaging speed multiphoton imaging. This improves the robustness and practicality of the optical system and increases imaging efficiency. Therefore, it solves the problems in related technologies, such as the need to measure the imaging environment before acquiring the distorted phase, which leads to a decrease in system imaging speed, and the reliance on active optical devices in the wavefront distortion compensation process, which increases the complexity of the optical system, makes it impossible to quickly achieve high-quality multiphoton imaging, reduces the imaging efficiency of the optical system, and results in low practicality.

[0080] Figure 9 A schematic diagram of the structure of an electronic device provided in an embodiment of this application. The electronic device may include:

[0081] The memory 901, the processor 902, and the computer program stored on the memory 901 and capable of running on the processor 902.

[0082] When the processor 902 executes the program, it implements the digital adaptive optics method for multiphoton imaging provided in the above embodiments.

[0083] Furthermore, electronic devices also include:

[0084] Communication interface 903 is used for communication between memory 901 and processor 902.

[0085] The memory 901 is used to store computer programs that can run on the processor 902.

[0086] The memory 901 may include high-speed RAM memory, and may also include non-volatile memory, such as at least one disk storage device.

[0087] If the memory 901, processor 902, and communication interface 903 are implemented independently, then the communication interface 903, memory 901, and processor 902 can be interconnected via a bus to complete communication between them. The bus can be an Industry Standard Architecture (ISA) bus, a Peripheral Component Interconnect (PCI) bus, or an Extended Industry Standard Architecture (EISA) bus, etc. The bus can be divided into address bus, data bus, control bus, etc. For ease of representation, Figure 9The bus is represented by a single thick line, but this does not mean that there is only one bus or one type of bus.

[0088] Optionally, in a specific implementation, if the memory 901, processor 902, and communication interface 903 are integrated on a single chip, then the memory 901, processor 902, and communication interface 903 can communicate with each other through an internal interface.

[0089] The processor 902 may be a central processing unit (CPU), an application specific integrated circuit (ASIC), or one or more integrated circuits configured to implement the embodiments of this application.

[0090] This embodiment also provides a computer-readable storage medium storing a computer program that, when executed by a processor, implements the digital adaptive optics method for multiphoton imaging as described above.

[0091] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0092] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this application, "N" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0093] Any process or method described in the flowchart or otherwise herein can be understood as representing a module, segment, or portion of code comprising one or N executable instructions for implementing custom logic functions or processes, and the scope of the preferred embodiments of this application includes additional implementations in which functions may be performed not in the order shown or discussed, including substantially simultaneously or in reverse order depending on the functions involved, as should be understood by those skilled in the art to which embodiments of this application pertain.

[0094] The logic and / or steps represented in the flowchart or otherwise described herein, for example, can be considered as a sequenced list of executable instructions for implementing logical functions, and can be embodied in any computer-readable medium for use by, or in conjunction with, an instruction execution system, apparatus, or device (such as a computer-based system, a processor-included system, or other system that can fetch and execute instructions from, an instruction execution system, apparatus, or device). For the purposes of this specification, "computer-readable medium" can be any means that can contain, store, communicate, propagate, or transmit programs for use by, or in conjunction with, an instruction execution system, apparatus, or device. More specific examples (a non-exhaustive list) of computer-readable media include: an electrical connection having one or more wires (electronic device), a portable computer disk drive (magnetic device), random access memory (RAM), read-only memory (ROM), erasable and editable read-only memory (EPROM or flash memory), fiber optic devices, and portable optical disc read-only memory (CDROM). Alternatively, the computer-readable medium may be paper or other suitable media on which the program can be printed, since the program can be obtained electronically by optically scanning the paper or other medium, followed by editing, interpreting, or otherwise processing as necessary, and then stored in a computer memory.

[0095] It should be understood that the various parts of this application can be implemented using hardware, software, firmware, or a combination thereof. In the above embodiments, the N steps or methods can be implemented using software or firmware stored in memory and executed by a suitable instruction execution system. For example, if implemented in hardware as in another embodiment, it can be implemented using any one or a combination of the following techniques known in the art: discrete logic circuits having logic gates for implementing logical functions on data signals, application-specific integrated circuits (ASICs) having suitable combinational logic gates, programmable gate arrays (PGAs), field-programmable gate arrays (FPGAs), etc.

[0096] Those skilled in the art will understand that all or part of the steps of the methods in the above embodiments can be implemented by a program instructing related hardware. The program can be stored in a computer-readable storage medium, and when executed, the program includes one or a combination of the steps of the method embodiments.

[0097] Furthermore, the functional units in the various embodiments of this application can be integrated into a processing module, or each unit can exist physically separately, or two or more units can be integrated into a module. The integrated module can be implemented in hardware or as a software functional module. If the integrated module is implemented as a software functional module and sold or used as an independent product, it can also be stored in a computer-readable storage medium.

[0098] The storage medium mentioned above can be a read-only memory, a disk, or an optical disk, etc. Although embodiments of this application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting this application. Those skilled in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of this application.

Claims

1. A digital adaptive optics method for multiphoton imaging, characterized in that, Includes the following steps: Based on two-photon synthetic aperture microscope, a small aperture excitation beam constrained by a pre-set diffraction-limited small aperture is used to collect three-dimensional sample projections from different perspectives and generate small aperture imaging results from different perspectives. Based on the small aperture imaging results, the lateral offset vectors generated by wavefront distortion phase caused by system distortion or tissue scattering during the imaging process are calculated, and three-dimensional sample information that meets the preset resolution conditions is obtained. as well as In the process of obtaining the three-dimensional reconstruction result based on the three-dimensional sample information, optical distortion is compensated according to the lateral offset vector.

2. The method according to claim 1, characterized in that, In the process of obtaining the 3D reconstruction result based on the 3D sample information, compensating for optical distortion according to the lateral offset vector includes: The wavefront distortion phase determined by the lateral offset vector is applied to the generation of the system point spread function in the preset three-dimensional reconstruction algorithm.

3. The method according to claim 2, characterized in that, The derivative function of the lateral offset vector and the wavefront distortion phase satisfies the following relationship: Where Δφ is the amount of distortion phase change, and φ(x,y) is the distorted incident light wavefront function. The offset vector on the X-axis of the image for each small aperture image. This is the offset vector on the Y-axis of the image for each small aperture image.

4. The method according to claim 2, characterized in that, In the process of obtaining the 3D reconstruction result based on the 3D sample information, compensating for optical distortion according to the lateral offset vector includes: In the preset 3D reconstruction algorithm, the lateral offset vector is compensated before 3D reconstruction is performed.

5. A digital adaptive optics device for multiphoton imaging, characterized in that, include: The acquisition module is used to acquire three-dimensional sample projections from different perspectives based on a two-photon synthetic aperture microscope. It utilizes a small aperture excitation beam constrained by a preset diffraction-limited aperture to generate small aperture imaging results from different perspectives. The calculation module is used to calculate the lateral offset vector of the image with different sub-apertures caused by wavefront distortion phase caused by system distortion or tissue scattering during the imaging process, based on the small aperture imaging results, and to obtain three-dimensional sample information that meets the preset resolution conditions. as well as The imaging module is used to compensate for optical distortion according to the lateral offset vector during the process of obtaining the three-dimensional reconstruction result based on the three-dimensional sample information.

6. The apparatus according to claim 5, characterized in that, The imaging module includes: The correction unit is used to apply the wavefront distortion phase determined by the lateral offset vector to the generation of the system point spread function in the preset three-dimensional reconstruction algorithm.

7. The apparatus according to claim 6, characterized in that, The derivative function of the lateral offset vector and the wavefront distortion phase satisfies the following relationship: Where Δφ is the amount of distortion phase change, and φ(x,y) is the distorted incident light wavefront function. The offset vector on the X-axis of the image for each small aperture image. This is the offset vector on the Y-axis of the image for each small aperture image.

8. The apparatus according to claim 6, characterized in that, The imaging module includes: The compensation unit is used to compensate the lateral offset vector in the preset 3D reconstruction algorithm before performing 3D reconstruction.

9. An electronic device, characterized in that, include: A memory, a processor, and a computer program stored in the memory and executable on the processor, the processor executing the program to implement the digital adaptive optics method for multiphoton imaging as described in any one of claims 1-4.

10. A computer-readable storage medium having a computer program stored thereon, characterized in that, The program is executed by the processor to implement the digital adaptive optics method for multiphoton imaging as described in any one of claims 1-4.

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