Image processing method and apparatus for arbitrary phase modulation microscopy

Abstract

The application provides an image processing method and device for a microscope under arbitrary phase modulation. An amplitude mask is configured in a microscopic imaging system. A first intensity image of a sample collected by the microscopic imaging system is obtained. After a phase optical transfer function of the system is obtained, effective transfer functions composed of a plurality of preset phase values and the phase optical transfer function are used to pre-process the first intensity image and perform deconvolution operation. While obtaining a target image of the sample with high contrast and resolution, a target phase shift introduced by the system configuring the amplitude mask is determined and stored in association with the system. The target phase shift can be directly called to perform deconvolution operation on other intensity images collected by the same microscopic imaging system, and to reconstruct phase information or refractive index information of the sample. The image processing efficiency is improved, and the image processing method and device are low in cost and easy to implement.

Description

Technical Field

[0001] This application relates to the field of microscopy, specifically to an image processing method and device for a microscope under arbitrary phase modulation. Background Technology

[0002] Phase contrast microscopy, with its non-contact and non-invasive label-free imaging of transparent samples, is an effective tool for long-term dynamic observation of transparent living biological samples (such as cells). In practical applications, traditional phase contrast microscopy techniques typically place a phase ring with specific phase modulation (such as ±0.5π) and amplitude attenuation at the rear pupil plane (a specially designed phase contrast objective) or the conjugate plane of the rear pupil plane of the phase contrast microscope. This improves the contrast of label-free imaging of transparent samples and promotes the development of label-free microscopy techniques.

[0003] However, traditional phase-contrast microscopy relies on phase-contrast objectives, which are costly and require the numerical aperture of the illumination to match the phase-contrast ring on the objective. This affects the resolution of phase-contrast microscopy and the collection of weak fluorescence signals, thus limiting the application range of phase-contrast microscopy. Summary of the Invention

[0004] To address the aforementioned technical problems, this application provides the following technical solutions:

[0005] This application proposes an image processing method for microscopes under arbitrary phase modulation, the method comprising:

[0006] The first intensity image and phase optical transfer function of the sample are obtained under a microscopic imaging system; the first intensity image is acquired after a frame mask is configured in the microscopic imaging system.

[0007] The first intensity image is preprocessed to obtain the second intensity image;

[0008] An effective transfer function is obtained by using multiple preset phase values ​​and the phase optical transfer function;

[0009] Using the effective transfer function, the second intensity image is deconvolved to obtain the target image of the sample, as well as the target phase shift generated by the amplitude mask introduced by the microscopic imaging system;

[0010] The target phase shift is associated with the system identifier of the microscopic imaging system and stored, so as to reconstruct images of other intensities acquired by the microscopic imaging system using the target phase shift.

[0011] Optionally, the preprocessing of the first intensity image to obtain the second intensity image includes:

[0012] The first intensity image is subjected to background removal processing to obtain a background-removed intensity image;

[0013] The background-removed intensity image is subjected to background normalization processing to obtain a second intensity image.

[0014] Optionally, the step of performing a deconvolution operation on the second intensity image using the effective transfer function to obtain the target phase shift generated by the amplitude mask configured in the microscopic imaging system includes:

[0015] The effective transfer function is deconvolved with the second intensity image to obtain the deconvolution results corresponding to each of the plurality of preset phase values; the deconvolution results can characterize the image contrast of the corresponding reconstructed image.

[0016] Compare multiple deconvolution results and select a preset phase value corresponding to the deconvolution result that represents the highest image contrast.

[0017] The selected preset phase value is determined as the target phase shift generated by configuring the amplitude mask in the microscopic imaging system.

[0018] Optionally, during the deconvolution operation of the second intensity image using the effective transfer function, the incident light on the sample under the microscopic imaging system is amplitude modulated to reconstruct the phase information or refractive index information of the sample; the amplitude modulation can cause any phase shift of the target to be determined.

[0019] Optionally, the phase optical transfer function includes a first optical transfer function corresponding to a first point diffusion function and a second optical transfer function corresponding to a second point diffusion function; the first point diffusion function and the second point diffusion function are respectively used to represent the relationship between the conjugate function corresponding to the second intensity image and the sample scattering;

[0020] The process of obtaining an effective transfer function by utilizing multiple preset phase differences and the phase optical transfer function includes:

[0021] Multiple preset phase values ​​can be obtained;

[0022] An effective transfer function is generated using the multiple preset phase values, the first optical transfer function, and the second optical transfer function.

[0023] Optionally, the step of performing a deconvolution operation on the second intensity image using the effective transfer function includes:

[0024] Based on the transformation relationship between the sample scattering potential, the point spread function, and the conjugate function of the second intensity image, the expression of the second intensity image is adjusted; the adjusted expression of the second intensity image can represent the convolution relationship between the point spread function and the sample scattering potential.

[0025] The second intensity image is deconvolved using the effective transfer function.

[0026] Optionally, the method further includes:

[0027] Obtain the intensity image of the sample to be processed acquired under the microscopic imaging system, and retrieve the target phase shift associated with the system identifier of the microscopic imaging system;

[0028] Based on the target phase shift, the preprocessed intensity image to be processed is reconstructed to obtain the target image of the sample being tested.

[0029] This application also proposes an image processing apparatus for a microscope under arbitrary phase modulation, the apparatus comprising:

[0030] The data acquisition module is used to acquire the first intensity image and phase optical transfer function of the sample under the microscopic imaging system; the first intensity image is acquired after a frame mask is introduced into the microscopic imaging system.

[0031] The preprocessing module is used to preprocess the first intensity image to obtain the second intensity image;

[0032] An effective transfer function acquisition module is used to obtain an effective transfer function using multiple preset phase differences and the phase optical transfer function;

[0033] The deconvolution operation module is used to perform a deconvolution operation on the second intensity image using the effective transfer function to obtain the target image of the sample and the target phase difference generated by the amplitude mask introduced by the microscopic imaging system.

[0034] The data storage module is used to associate the target phase difference with the system identifier of the microscopic imaging system and store it, so as to use the target phase difference to reconstruct images of other intensities acquired by the microscopic imaging system.

[0035] This application also proposes a computer device, the computer device comprising:

[0036] Communication module;

[0037] At least one memory is used to store a program that implements the image processing method for a microscope under arbitrary phase modulation as described above;

[0038] At least one processor is configured to load and execute the program stored in the memory to implement the above-described image processing method for a microscope under arbitrary phase modulation.

[0039] This application also proposes a computer-readable storage medium, characterized in that it stores a plurality of computer instructions, which are loaded and executed by a processor to implement the image processing method for a microscope under arbitrary phase modulation as described above.

[0040] Based on the above technical solution, this application proposes an image processing method and device for microscopes under arbitrary phase modulation. To reduce costs, ensure high image contrast, and expand the scope of application, a amplitude mask is configured in the microscopic imaging system. After acquiring the first intensity image of the sample and the phase optical transfer function of the system under the microscopic imaging system, in order to determine the phase shift introduced by the amplitude mask, this application uses an effective transfer function composed of multiple preset phase values ​​and the phase optical transfer function to perform a deconvolution operation on the preprocessed intensity image. While acquiring a high-contrast and high-resolution target image of the sample, the target phase shift introduced by the amplitude mask of the system is determined and stored in association with the system. Subsequently, the target phase shift can be directly called. Other intensity images acquired under the same microscopic imaging system are preprocessed using the same steps and then deconvolved to reconstruct the phase information or refractive index information of the sample, thereby meeting the requirements of high resolution and high contrast while improving image processing efficiency. Attached Figure Description

[0041] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of this application. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0042] Figure 1 This is a schematic flowchart of an optional example of the image processing method for a microscope under arbitrary phase modulation proposed in this application;

[0043] Figure 2 This is a flowchart illustrating yet another optional example of the image processing method for a microscope under arbitrary phase modulation proposed in this application.

[0044] Figure 3a This is a schematic diagram of the original data and the results of different processing methods in Example 1 of the optional scenario of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0045] Figure 3b This is a schematic diagram of the processing results corresponding to four preset phase values ​​with high contrast in the original data and image, in Example 1 of the optional scenario of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0046] Figure 3c This is a schematic diagram of the normalized curves of the processing results obtained by different processing methods for the dashed line at the letter d in Example 1 of the optional scenario of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0047] Figure 3d This is a schematic diagram of the normalized curves of the processing results obtained by different processing methods for the dashed line at the letter 'e' in Example 1 of the optional scenario of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0048] Figure 4a This is a schematic diagram of the original data and the results of different processing methods in Example 2 of the optional scenario for the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0049] Figure 4b This is a schematic diagram of the processing results corresponding to four preset phase values ​​with high contrast in the original data and the image, in Example 2 of the optional scenario of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0050] Figure 4c This is a schematic diagram of the normalized curves of the processing results obtained by different processing methods for the dashed line at the letter d in Example 2 of the optional scenario of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0051] Figure 4d This is a schematic diagram of the normalized curves of the processing results obtained by different processing methods for the dashed line at the letter 'e' in Example 2 of the optional scenario of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0052] Figure 5a This is a schematic diagram of the original data and the results of different methods in Example 3 of the optional scenario for the image processing method for microscopes under arbitrary phase modulation proposed in this application;

[0053] Figure 5b This is a schematic diagram of the normalized curve of the processing result in Example 3 of the optional scenario for the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0054] Figure 6a This is a schematic diagram of the original data and the results of different methods in Example 4 of the optional scenario for the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0055] Figure 6b This is a schematic diagram of the normalized curve of the processing result in Example 4 of the optional scenario for the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0056] Figure 7 This is a schematic diagram of an optional example of the image processing apparatus for a microscope under arbitrary phase modulation proposed in this application;

[0057] Figure 8 This is a schematic diagram of another optional example of the image processing apparatus for a microscope under arbitrary phase modulation proposed in this application;

[0058] Figure 9 This is a schematic diagram of the hardware structure of an optional example of the computer device proposed in this application. Detailed Implementation

[0059] Regarding the description in the background section, a phase ring can be used to convert the phase change caused by the refractive index change of a transparent sample into an observable change in light intensity. By modulating the phase of the non-scattered light (i.e., incident light) that passes through the sample and does not interact with it, its phase can be increased or decreased by 0.5π, and the incident light can be attenuated by a certain amplitude, thereby improving the image contrast. However, this requires that the numerical aperture of the illumination must match the phase contrast ring on the objective lens. At the same time, the phase of the phase ring cannot be arbitrary, which has significant limitations and will affect the image resolution.

[0060] To address the aforementioned issues, a method is proposed to incorporate an amplitude attenuation mask into a conventional microscopic imaging system (such as a microscope using a non-contrast objective) to achieve phase contrast, based on experimental conditions and the required final image resolution. Specifically, an amplitude attenuation mask (introducing an unknown phase) is placed at the conjugate plane of the objective's pupil plane to meet the demands for image resolution and contrast. This mask can be fabricated using commercially available low-cost optical density attenuation sheets, making it easy to process. Furthermore, different mask sizes can be flexibly replaced to match the size of the illumination source, thus expanding the application range of phase-contrast microscopy.

[0061] However, the thickness of the coating on the glass plate used to manufacture the optical density attenuator is generally not precisely controllable. Therefore, arbitrary phase shifts can occur when incident light passes through the plate. When using the aforementioned amplitude attenuation mask to attenuate non-scattered light, the imaging contrast may be lower than that of a conventional phase-contrast microscope with a phase shift of ±0.5π. To address this, this application proposes a universal image reconstruction method for phase-contrast images with arbitrary phase modulation, enabling the acquisition of high-contrast images applicable to bright-field microscopes, phase-contrast microscopes, and optical systems that introduce arbitrary phase modulation with only amplitude attenuation.

[0062] Based on the easily understandable technical concept and technical requirements derived from the analysis of this application above, this application proposes to use deconvolution to determine the phase shift introduced by the system, thereby obtaining images with higher contrast. It does not require matching the numerical aperture of the illumination with the numerical aperture of the objective lens. Furthermore, by using a large numerical aperture objective lens and a high-resolution microscopic imaging system, it is possible to recover high-contrast information from a label-free transparent sample by acquiring a set of intensity images, thus having a greater advantage in achieving high-resolution label-free imaging.

[0063] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0064] Reference Figure 1 This is a flowchart illustrating an optional example of the image processing method for a microscope under arbitrary phase modulation proposed in this application. The method can be applied to computer equipment, which can be a server or a terminal. The server can be a physical server or a desktop server. The terminal can include microscopes with data processing capabilities, desktop computers, laptops, smart medical devices, etc. This application does not limit the product type of computer equipment or its application environment, and can be determined as appropriate.

[0065] like Figure 1 As shown, the image processing method for microscopes under arbitrary phase modulation proposed in this embodiment may include, but is not limited to, the following steps:

[0066] Step S11: Obtain the first intensity image and phase optical transfer function of the sample under the microscopic imaging system;

[0067] Based on the above description of the technical solution proposed in this application, a phase-contrast mask can be introduced into microscopic imaging systems such as bright-field microscopes and phase-contrast microscopes (which have non-contrast objectives) to achieve phase contrast, thereby improving the resolution and contrast of phase-contrast images under arbitrary phase modulation. Therefore, after placing a mask for amplitude attenuation (denoted as the amplitude mask) at the conjugate plane of the pupil plane of a conventional objective lens in the microscopic imaging system, the sample is placed in the scanned area, and the sample is scanned and acquired by the microscopic imaging system to obtain the intensity image of the sample, i.e., the first intensity image acquired for the first time under this microscopic imaging system.

[0068] Subsequently, if the computer device is not a microscope, the acquired first intensity image can be sent to the computer device via wired or wireless means; if the computer device is a microscope, subsequent processing steps can be executed directly.

[0069] The expression for the first intensity image I acquired by the aforementioned microscopic imaging system can be:

[0070]

[0071] In the above first intensity image expression (1), a can represent the amplitude attenuation achieved by the amplitude mask added to the above microscopic imaging system, φ can represent the phase difference (i.e., unknown phase shift) introduced by the mask, u i u s These can represent the incident light field distribution and the scattered light field distribution, respectively. It should be noted that, for the scenario where the sample is a weakly scattering transparent sample, the expression for the first intensity image obtained... This can be ignored; therefore, the above formula (1) can be transformed into:

[0072]

[0073] In the above formula (2), It can represent a background image. and The terms are conjugates and can be called conjugate functions. This application does not limit the content of the expression for the intensity image.

[0074] The implementation of the Phase Optical Transfer Function (POTF) obtained above can be obtained through theoretical calculations, and this application will not elaborate on the calculation method of POTF. It should be noted that the initial phase difference (phase shift) in this phase optical transfer function can be zero, denoted as φ. 初始 =0.

[0075] Step S12: Preprocess the first intensity image to obtain the second intensity image;

[0076] To improve image processing efficiency and accuracy, this application first removes interference information, such as background images unrelated to the sample, from the directly acquired first intensity image. Therefore, this embodiment performs background removal processing on the first intensity image, and this application does not limit the specific implementation method.

[0077] Subsequently, to achieve quantization uniformity of the parameters in the intensity image expression, normalization processing can be performed. For example, background normalization processing can be performed on the first intensity image after background removal, and the preprocessed intensity image can be denoted as the second intensity image. It should be noted that the preprocessing method for the first intensity image, including but not limited to the background removal and normalization processing methods described above, can be determined according to actual needs.

[0078] Step S13: Using multiple preset phase values ​​and the phase optical transfer function, obtain the effective transfer function;

[0079] To perform the deconvolution operation on the aforementioned second-intensity image and determine the unknown phase shift (phase difference) introduced by the amplitude mask in the microscopic imaging system, embodiments of this application propose using a series of known different phase values ​​to calculate different effective transfer functions to verify the accuracy of the image processing method proposed in this application. In this verification process, this known phase-modulated data can be processed as unknown phase data, and a series of preset phase values ​​can be used to predict the value of the unknown phase shift introduced by the amplitude mask.

[0080] The aforementioned series of preset phase values ​​can be obtained by methods such as interferometric detection, or by prediction based on the first intensity image acquired by the aforementioned microscopic imaging system. This application does not limit the preset implementation method for obtaining multiple preset phase φ values, nor the specific values ​​of these multiple φ values, and can be determined as appropriate.

[0081] Step S14: Using the effective transfer function, perform deconvolution operation on the second intensity image to obtain the target image of the sample and the target phase shift generated by the amplitude mask configured in the microscopic imaging system.

[0082] In this embodiment, a series of known preset phase values ​​are used to calculate different effective transfer functions and deconvolve them with the preprocessed intensity image (the second intensity image mentioned above). The phase shift introduced by the system is determined by the contrast of the deconvolution result. The higher the deconvolution contrast, the closer the preset phase difference is to the phase shift introduced by the system.

[0083] Based on this, when the contrast of the above deconvolution results is very close, the approximate range of the phase shift introduced by the microscopic imaging system can be initially locked. By performing deconvolution operation on each phase value within this approximate range, the contrast of the recovered images may be very close. This application can arbitrarily select one phase value as the target phase shift to improve the contrast of the intensity image subsequently acquired by the microscopic imaging system.

[0084] It should be noted that this application does not describe in detail how to implement the deconvolution operation on the effective transfer function and the second intensity image. The deconvolution algorithm used can be, but is not limited to, the above step S14.

[0085] Step S15: The target phase shift is associated with the system identifier of the microscopic imaging system and stored, so as to use the target phase shift to reconstruct images of other intensities acquired by the microscopic imaging system.

[0086] In this application, for the same microscopic imaging system, the approximate range of phase shift caused by the addition of an amplitude mask to the microscopic imaging system is determined. That is, the above-mentioned operation method of locking the approximate range of phase shift introduced by the microscopic imaging system can be performed once. For other intensity images acquired subsequently, the target phase shift determined this time can be used for image reconstruction processing to improve the contrast of images acquired by microscopic imaging systems such as bright-field microscopes and phase-contrast microscopes with arbitrary phase modulation.

[0087] Furthermore, compared to spatial interferometry, which requires acquiring multiple sets of intensity images with different phase modulations to recover the scattering potential information of the sample, the reconstruction method described above in this application only requires acquiring one set of intensity images to recover the refractive index and phase information of the sample. Especially in applications using large numerical aperture objectives and high-resolution microscopic imaging systems, high-contrast information can be recovered from unmarked transparent samples using only one set of intensity images. Moreover, in this process, only the amplitude of the unscattered light of the microscopic imaging system can be modulated, improving image processing efficiency and application range.

[0088] Reference Figure 2 This is a flowchart illustrating another optional example of the image processing method for microscopes under arbitrary phase modulation proposed in this application. This embodiment describes an optional refined implementation of the image processing method for microscopes under arbitrary phase modulation described above, but is not limited to the refined implementation method described in this embodiment. This refined implementation method can still be executed by a computer device, such as... Figure 2 As shown, the method may include:

[0089] Step S21: Obtain the first intensity image of the sample under the microscopic imaging system;

[0090] Step S22: Perform background removal processing on the first intensity image to obtain a background-removed intensity image;

[0091] Step S23: Perform background normalization processing on the intensity image after background removal to obtain the second intensity image;

[0092] In this embodiment of the application, in conjunction with the above description of the expression of the first intensity image, in order to achieve deconvolution of the preprocessed intensity image, the conjugate image information of the preprocessed intensity image can be analyzed first. Therefore, this application can use the phase optical point spread function (which can be simply referred to as the point spread function) to transform the conjugate function in the expression of the first intensity image, and record the intensity image of the obtained expression as the second intensity image. The implementation process is not limited in this application.

[0093] Regarding the expression for the first intensity image shown in formula (2) above, the background image in the first intensity image can be removed first, i.e., interference information can be eliminated to improve the efficiency and accuracy of subsequent image processing. Optionally, during the background removal process of the first intensity image, the mean value of the first intensity image can be used to approximate its background image. Therefore, in this embodiment, the background image to be removed can be a constant value, denoted as I. B Based on the above description of the expression for the first intensity image, Therefore, after removing the background from the first intensity image, the expression for the resulting intensity image after background removal can be:

[0094]

[0095] In practical applications, factors such as the image acquisition device itself, the acquisition environment, and imaging factors may cause inconsistencies in the grayscale information of the same tissue in the intensity image. This application can use image normalization processing to reduce or even eliminate grayscale inconsistencies in the image while retaining valuable grayscale differences. Therefore, this application can perform background normalization processing on the background-removed intensity image as shown in formula (3) to convert the intensity image into a standard form of the second intensity image I'. The expression of the second intensity image can be:

[0096]

[0097] In the above formula (4), combined with the relevant description of the expression of the intensity image before transformation above, and Terms that are conjugates of each other can be called conjugate functions, and and The spectrum of the conjugate function is antisymmetric in the fz direction. The expression of the conjugate function is not described in detail in this application.

[0098] Analysis revealed that the conjugate function in the intensity image expression can be represented as: the sample scattering potential v and the point spread function of the microscopic imaging system (such as h). 12 h 21 The result is obtained by convolution, such as Therefore, in this embodiment, the above formula (4) can be transformed to obtain the deformed expression of the second intensity image as shown in the following formula (5):

[0099]

[0100] In the above formula (5), a can represent the amplitude attenuation achieved by the amplitude mask configured in the microscopic imaging system, φ can represent the phase difference (also known as phase shift) introduced by the amplitude mask, v can represent the scattering potential of the sample, and h 12 It can represent The phase optical point spread function corresponding to this function is denoted as the first point spread function; h 21 It can represent The phase optical point spread function corresponding to this function is denoted as the second point spread function. Combining this with the description of the two conjugate terms above, we know that h... 12 and h 21 The corresponding spectrum is symmetrically distributed in the fz direction and has opposite signs.

[0101] In practical applications, the light vibration of any small surface element on the object plane can be diffused on the image plane (the conjugate plane of the object plane) after passing through a microscopic imaging system, generating a certain light field distribution, that is, the image field distribution function is called the point spread function. Based on this, the first point spread function and the second point spread function mentioned above in this application can be the point image object light field distribution function, and the method of obtaining them will not be described in detail.

[0102] It should be noted that, theoretically, this application can perform an inverse Fourier transform on the optical transfer function to obtain the corresponding point spread function, which represents the relationship between the conjugate function and the sample scattering potential. However, in the subsequent deconvolution operation, the transfer function is actually used for calculation, and the point spread function does not need to be calculated. Therefore, the point spread function in the deformed expression of the intensity image above does not need to be calculated.

[0103] Step S24: Using multiple preset phase values, the first optical transfer function, and the second optical transfer function, generate an effective transfer function;

[0104] In this embodiment of the application, the phase optical transfer function (POTF) analytical method can be used to theoretically calculate the first point diffusion function h. 12 The corresponding first optical transfer function H 12 , and the second point diffusion function h 21 The corresponding second optical transfer function H 21 The implementation process is not detailed in the embodiments of this application.

[0105] The optical transfer function (OPF) can be used to characterize the performance of a microscopic imaging system, such as its spectral transfer characteristics. Therefore, the phase optical transfer function proposed in this application can describe the modulation applied to each frequency component by the microscopic imaging system. This application does not elaborate on the contents of the aforementioned optical transfer functions. In some other embodiments proposed in this application, regarding the aforementioned H... 12 and H 21 The phase optical transfer function of the microscopic imaging system, as described above, can be obtained through theoretical calculations of the phase optical transfer function or through experimental measurements. This application addresses H... 12 and H 21 The actual method of obtaining it will not be detailed.

[0106] Based on the analysis above regarding the expression of the intensity image acquired by the microscopic imaging system, it can be seen that the intensity image of a sample can be obtained by convolving the sample information (such as unknown quantities like the sample's scattering potential) with the point spread function of the microscopic imaging system. For example, if the sample is an ideal point, it can become a spot on the image plane after passing through the microscopic imaging system; complex samples can be regarded as the superposition of countless points, and the acquired intensity image can be the superposition of countless spots. Therefore, during 3D sample imaging, the defocus information will enter the focal plane, resulting in a significant reduction in the contrast of the obtained image. After obtaining the point spread function of the microscopic imaging system according to the method described above, reverse engineering can be used to restore the defocus information as much as possible through deconvolution, thereby improving the image contrast.

[0107] Based on this, in order to perform a deconvolution operation on the preprocessed intensity image, combined with the intensity image expression described above, such as in formula (5) (ae -iφ h 12 +ae iφ h 21 The formula for H can be calculated using a set of phases with different phase values ​​φ, combined with the system transfer function, to obtain the effective transfer function, i.e., H = ae^(-φ / φ). -iφ H 12 +ae iφ H 21The method for obtaining these multiple preset phase values ​​φ can be referred to, but is not limited to, the description in the corresponding part of the above embodiment. It can also be obtained by random generation.

[0108] Optionally, this application may randomly select a series of preset phase values ​​from a preset phase range, such as [0,2π], [-π,π], etc. In other words, the above-mentioned multiple preset phase values ​​may be random values ​​within the preset phase range, and this application does not impose any restrictions on the value of each random value.

[0109] Step S25: Using the effective transfer function, perform deconvolution operation on the second intensity image to obtain the deconvolution results corresponding to each of the multiple preset phase values;

[0110] Based on the above description of the expression and deformation of the preprocessed intensity image, it can be seen that the expression of the adjusted second intensity image can represent the convolution relationship between the point spread function and the sample scattering potential. Specifically, the conjugate function of the second intensity image can be expressed as the convolution of the sample scattering potential and the point spread function. The point spread function can be obtained by the inverse Fourier transform of the corresponding optical transfer function. Therefore, the effective transfer function can be used to perform deconvolution operation on the preprocessed intensity image to obtain the scattering potential of the sample.

[0111] Step S26: Compare the multiple deconvolution results obtained, and select the preset phase value corresponding to the deconvolution result that represents the highest image contrast;

[0112] Step S27: The selected preset phase value is determined as the target phase shift generated by the amplitude mask configured in the microscopic imaging system;

[0113] Following the analysis above, since the closer the known preset phase value is to the phase shift introduced by the microscopic imaging system, the closer the effective transfer function is to the true optical transfer function of the system, and the higher the image contrast obtained by deconvolution. Based on this, this application can use a deconvolution operation to obtain deconvolution results for different preset phase values. After comparison, the preset phase value corresponding to the highest image contrast is determined, which is the phase value closest to the phase shift that causes the microscopic imaging system. In this application embodiment, the preset phase value can be directly determined as the unknown phase shift introduced by the addition of an amplitude mask for amplitude attenuation in the microscopic imaging system. Of course, this application can adaptively adjust the determined preset phase value and determine the adjusted phase value as the unknown phase shift introduced by the amplitude mask.

[0114] It should be noted that the implementation process of obtaining high-resolution and high-contrast phase-contrast images of samples using a microscopic imaging system with an amplitude mask is not described in detail in the embodiments of this application.

[0115] Step S28: The target phase shift is associated with the system identifier of the microscopic imaging system and stored, so as to use the target phase shift to reconstruct images of other intensities acquired by the microscopic imaging system.

[0116] For the aforementioned microscopic imaging system of this application, it is only necessary to execute the method described above once and record the target phase shift introduced by the amplitude mask in this calculation. When using the microscopic imaging system for image processing in the future, the target phase shift calculated in the first calculation can be directly retrieved and deconvolution operation can be performed in combination with the phase optical transfer function of the system to improve the contrast of the acquired intensity image and obtain the phase information and refractive index information of the sample.

[0117] Based on this, after obtaining the target phase shift introduced by the amplitude mask in the aforementioned microscopic imaging system, the target phase shift can be associated with and stored in the microscopic imaging system. In this way, without changing the scenario of the microscopic imaging system, the image of the sample under test can be acquired through the microscopic imaging system, and the acquired intensity image of the sample to be processed can be sent to a computer device, such as the processor or server or other terminal configured in the microscope. The computer device obtains the intensity image of the sample to be processed acquired by the microscopic imaging system, and can retrieve the target phase shift associated with the system identifier of the microscopic imaging system. Based on the target phase shift, the intensity image to be processed is reconstructed to obtain the target image of the sample under test. That is, according to the method described above, the target phase shift and the phase optical transfer function of the system are used to perform a deconvolution operation on the preprocessed intensity image to be processed to improve the contrast of the intensity image to be processed, so as to accurately and reliably obtain the reflectivity information and phase information of the sample under test.

[0118] It should be understood that in the case of a microscope where the computer equipment is the microscopic imaging system, the target phase shift can be directly stored. In this way, during subsequent image processing, the stored target phase shift can be directly read, improving the contrast of phase-contrast images under arbitrary phase modulation and meeting practical needs.

[0119] Based on the foregoing description of the image processing method for microscopes under arbitrary phase modulation proposed in this application, the following will provide comparative explanations with examples, but are not limited to the following examples:

[0120] Example 1 compares the processing results obtained by different image processing methods on the original data (intensity image) of a 200nm sphere under 0π modulation phase acquired by a spatial light interference microscope (SLIM) with phase modulation, to illustrate the improvement of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0121] For example Figure 3a The raw data of the sample acquired under the 0π modulation phase (z = 0 μm) shown in the first row of the attached figures can be processed using the image processing method for microscopes under arbitrary phase modulation proposed in this application. The processing procedure can be referred to the description of the method embodiment above. Using different preset phase values ​​φ and the system optical transfer function, a deconvolution operation is performed on the preprocessed raw data (such as background removal and normalization of the raw data) to obtain the following results: Figure 3a The second row of attached figures shows the deconvolution results for different preset phase values ​​φ, thus characterizing the recovered sample scattering potential. Then, the deconvolution results for four preset phase values ​​with higher contrast (e.g., φ = -0.4π, φ = -0.2π, φ = 0π, φ = 0.2π, each in its own...) can be selected. Figure 3a The original data (as shown in the attached figure) and the deconvolution results (denoted as SLIM-decon) of the four sets of phase modulation data acquired using SLIM are compared. Figure 3b As shown, this is used to verify the correctness of the target phase shift and reconstruction results determined in this application.

[0122] pass Figure 3b Comparative analysis shows that the image displayed on the xy-section of the magnified local image of the small ball has the highest contrast when φ = 0π, and the processing result of its xz-section is close to the result obtained by deconvolution using four sets of phase modulation data (such as the SLIM-decon mentioned above). Furthermore, regarding... Figure 3b The dashed lines corresponding to the letters 'd' and 'e' in Chinese are normalized using different processing methods to obtain the corresponding results, such as... Figure 3c The normalized curve shown in Figure 3d further proves that the processing result at φ = 0π is closest to the processing result of SLIM-decon. This shows that the target phase shift determined by the method described above (i.e., 0π obtained in this example) is consistent with the actual value, thus proving the correctness of the image processing method for microscopes under arbitrary phase modulation proposed in this application.

[0123] Example 2 compares the processed images (intensity images) of a 200nm sphere acquired under π-modulated phase using a spatial light interference microscopy (SLIM) with different image processing methods to illustrate the improvement of the image processing method proposed in this application for microscopy under arbitrary phase modulation. For example, Figure 4a The original data corresponding to the boxes in the first row of the attached figures, according to the image processing method described above in this application, is preprocessed using multiple preset phase values ​​and the system optical transfer function, and then deconvolution is performed to obtain the following: Figure 4a The second row shows various deconvolution processing results. You can then select the processing results corresponding to four preset phase values ​​with relatively high image contrast, the original data, and the SLIM-decon processing results for comparison and display, such as... Figure 4b As shown, the target phase shift (such as π in this example) determined by the processing method of this application is verified.

[0124] Based on the description of the corresponding part in Example 1, analyze Figure 4b The image with the highest contrast in the xy and xz sections of the magnified local image of the small and medium-sized spheres, with a preset phase value φ = π, and its xz section is also close to the SLIM-decon processing result. Additionally, as... Figure 4c and Figure 4d The corresponding letters shown (e.g.) Figure 4b The normalized curves of the different processing results of the dashed lines at the letters d and e in the figure are shown. The processing result under π phase modulation is closest to the SLIM-decon processing result, which once again proves the accuracy and reliability of the target phase shift determined by the above processing method proposed in this application. The contrast of the horizontal and vertical images of the above original data and reconstruction results has been significantly improved.

[0125] Example 3 compares and analyzes the raw data of a 20nm microsphere acquired by configuring an amplitude mask modulation for amplitude attenuation in a microscopic imaging system, and the corresponding processed results obtained by the image processing method for microscopes under arbitrary phase modulation proposed in this application. Figure 5a As shown in the first attached figure, this microscopic imaging system magnifies the image of the bounding area of ​​the original data (z = 0 μm) in the original tomographic data, resulting in the image shown below. Figure 5a The second row of attached figures shows an enlarged view of the original data, and the zx cross-sectional image corresponding to the dashed line within the box is obtained. Then, the processing method proposed in this application can be used to obtain the processing results corresponding to multiple preset phase values, including but not limited to... Figure 5aThe diagram shows several phase values. Similarly, further phase values ​​can be obtained. Figure 5a The letters 'c' and 'd' correspond to the normalized curves of different processing methods shown in the dashed lines, as follows: Figure 5b As shown, this demonstrates the accuracy and reliability of the image processing method proposed in this application.

[0126] Example 4: Raw COS7 cell data acquired using mask modulation with amplitude attenuation, and processed according to the method proposed in this application, yields the following result: Figure 6a The processing results shown are for Figure 6a The boxed area in the first row of the original data chart is enlarged to obtain the following: Figure 6a The second row shows an enlarged view of the original data and the xz cross-sectional view corresponding to the dashed line. Multiple preset phase values ​​are selected, and deconvolution processing is performed according to the method proposed in this application to obtain the following... Figure 6a The processing results shown, upon comparison, are identical to the verification results in Example 3 above; both indicate that the image contrast is best under phase modulation with φ = 0.2π. This can also be seen from... Figure 6b The verification result can be obtained from the normalized curves of the different processing results of the dashed lines corresponding to the letters c and d.

[0127] It should be noted that the raw data for Examples 3 and 4 above came from the same microscopic imaging system configured with an amplitude mask for amplitude attenuation. Figure 5a In the magnified partial image of the small ball shown, the image contrast of the xy and zx sections corresponding to φ = 0π, 0.2π, 0.4π, and 0.6π is relatively close. Comparing the zx section, the processing result with φ = 0π can be ruled out. Through the above normalization curve analysis, the processing result with better image contrast and resolution, namely φ = 0.2π, can be identified. Similarly, Figure 6a and Figure 6b Similar patterns have been observed, but will not be detailed in this embodiment.

[0128] Therefore, it can be inferred that the amplitude mask introduces a phase shift of approximately 0.2π while performing amplitude attenuation. This also proves that for the same microscopic imaging system, the operation of determining the introduced phase shift only needs to be performed once. Changing the sample under test will not cause a phase shift in the system. A set of data can be used for phase calculation, and the target phase shift obtained from the first calculation can be directly used for deconvolution processing, which greatly reduces the amount of computation and processing efficiency.

[0129] Reference Figure 7This is a schematic diagram of an optional example of an image processing apparatus for a microscope under arbitrary phase modulation proposed in this application. This apparatus can be applied to a microscopic imaging system incorporating an amplitude mask for amplitude attenuation. This application does not limit the type of system or its operating equipment; it can be determined as appropriate. Figure 7 As shown, the device may include:

[0130] The data acquisition module 71 is used to acquire the first intensity image and phase optical transfer function of the sample under the microscopic imaging system; the first intensity image is acquired after a frame mask is introduced into the microscopic imaging system.

[0131] Preprocessing module 72 is used to preprocess the first intensity image to obtain a second intensity image;

[0132] The effective transfer function acquisition module 73 is used to obtain the effective transfer function using multiple preset phase differences and the phase optical transfer function;

[0133] The deconvolution operation module 74 is used to perform a deconvolution operation on the second intensity image using the effective transfer function to obtain the target image of the sample and the target phase difference generated by the amplitude mask introduced by the microscopic imaging system.

[0134] In the embodiments of this application, the amplitude of the incident light on the sample under the microscopic imaging system can be modulated so that the system can introduce arbitrary phase shift. Then, the phase shift introduced thereby, i.e. the target phase shift, can be determined according to the image processing method described in the context embodiments of this application, so as to reconstruct the phase information or refractive index information of the sample and meet the scanning requirements of the sample.

[0135] The data storage module 75 is used to associate the target phase difference with the system identifier of the microscopic imaging system and store it, so as to use the target phase difference to reconstruct images of other intensities acquired by the microscopic imaging system.

[0136] In some embodiments, such as Figure 8 As shown, the preprocessing module 72 may include:

[0137] Background removal unit 721 is used to perform background removal processing on the first intensity image to obtain a background-removed intensity image;

[0138] Background normalization processing unit 722 is used to perform background normalization processing on the intensity image after background removal to obtain a second intensity image;

[0139] In some other embodiments, such as Figure 8 As shown, the deconvolution operation module 74 described above may include:

[0140] The deconvolution operation unit 741 is used to perform a deconvolution operation on the effective transfer function and the second intensity image to obtain the deconvolution result corresponding to each of the plurality of preset phase values; the deconvolution result can characterize the image contrast of the corresponding reconstructed image.

[0141] Comparison unit 742 is used to compare multiple deconvolution results and select a preset phase value corresponding to the deconvolution result that represents the highest image contrast.

[0142] The target phase shift determination unit 743 is used to determine the selected preset phase value as the target phase shift generated by the configuration of the amplitude mask in the microscopic imaging system.

[0143] Optionally, in the case where the phase optical transfer function of the microscopic imaging system includes a first optical transfer function corresponding to the first point diffusion function and a second optical transfer function corresponding to the second point diffusion function, as analyzed above, the first point diffusion function and the second point diffusion function are respectively used to represent the relationship between the conjugate function corresponding to the second intensity image (such as the two conjugate terms in the above expression) and the scattering potential of the sample. Based on this, the effective transfer function acquisition module 73 may include:

[0144] The preset phase value acquisition unit is used to acquire multiple preset phase values; for example, multiple preset phase values ​​are determined randomly and then input into the value system so that the system can acquire these multiple preset phase values; or multiple preset phase values ​​can be randomly selected from the preset phase range after inputting into the system, but it is not limited to this method of acquiring preset phase values.

[0145] An effective transfer function generation unit is used to generate an effective transfer function using the plurality of preset phase values, the first optical transfer function, and the second optical transfer function.

[0146] Optionally, based on the above analysis, the deconvolution operation module 74 may also include:

[0147] An adjustment unit is used to adjust the expression of the second intensity image based on the transformation relationship between the sample scattering potential, the point spread function, and the conjugate function of the second intensity image; the adjusted expression of the second intensity image can represent the convolution relationship between the point spread function and the sample scattering potential.

[0148] The deconvolution unit is used to perform a deconvolution operation on the second intensity image using the effective transfer function.

[0149] Based on the description of the embodiments above, the apparatus may further include:

[0150] The intensity image acquisition module is used to acquire the intensity image of the sample to be processed acquired under the microscopic imaging system.

[0151] The target phase shift retrieval module is used to retrieve the target phase shift stored in association with the system identifier of the microscopic imaging system.

[0152] The image reconstruction module is used to reconstruct the preprocessed intensity image based on the target phase shift to obtain the target image of the tested sample. The implementation process can be combined with the deconvolution operation process described above, which will not be described in detail in this embodiment.

[0153] It should be noted that the various modules and units in the above-mentioned device embodiments can all be stored in the memory as program modules. The processor executes the above-mentioned program modules stored in the memory to realize the corresponding functions. The functions realized by each program module and its combination, as well as the technical effects achieved, can be referred to the description of the corresponding part of the above-mentioned method embodiments. This embodiment will not repeat them here.

[0154] This application also provides a computer-readable storage medium storing a computer program, which is loaded and executed by a processor to implement the steps of the above-described image processing method for a microscope under arbitrary phase modulation. The specific implementation process can be referred to the description of the corresponding part of the above embodiments, and will not be repeated in this embodiment.

[0155] This application also proposes a computer program product or computer program, which includes computer instructions stored in a computer-readable storage medium. A processor of a computer device reads the computer instructions from the computer-readable storage medium and executes the computer instructions, causing the computer device to perform the methods provided in various optional implementations of the image processing method for microscopes under arbitrary phase modulation described above. Specific implementation processes can be referred to the descriptions of the corresponding embodiments above, and will not be repeated here.

[0156] Reference Figure 9 This is a schematic diagram of the hardware structure of an optional example of the computer device proposed in this application. As described in the corresponding part of the above embodiments, the computer device can be a server or a terminal, or a server and a terminal working together to implement the image processing method for microscopes under arbitrary phase modulation proposed in this application, depending on the circumstances. Figure 9 As shown, taking a computer device as a server as an example, the computer device may include: a communication module 91, at least one memory 92, and at least one processor 93, wherein:

[0157] The communication module 91 may include a communication module capable of data interaction via a wireless communication network, such as a WIFI module, a 5G / 6G (fifth-generation mobile communication network / sixth-generation mobile communication network) module, a GPRS module, etc. For example, it can directly communicate with the microscope where the microscopic imaging system is located to receive the intensity image of the sample acquired by the microscope, or the microscope can send the intensity image of the acquired sample to a local terminal via wired or wireless means, and then send it to the server for further processing using the local terminal's communication system, and feed back the final processing result to a preset terminal for display, etc. It can be determined according to the needs of the scenario. This application does not limit the communication connection method between different devices.

[0158] In addition, the aforementioned communication module 91 may also include a communication interface for enabling data interaction between internal components of a computer device, such as a USB interface, serial / parallel port, I / O opening, etc. This application does not limit the specific contents included in the communication module 91.

[0159] The memory 92 can be used to store a program that implements the image processing method for a microscope under arbitrary phase modulation described in the above method embodiments; the processor 93 can load and execute the program stored in the memory to implement the various steps of the image processing method for a microscope under arbitrary phase modulation described in the above corresponding method embodiments. The specific implementation process can be referred to the description of the corresponding part of the above embodiments, and will not be repeated here.

[0160] In practical applications, the communication module 91, memory 92 and processor 93 can be connected to a communication bus to realize data interaction between themselves and other structural components of the computer device. The specific details can be determined according to actual needs, and will not be described in detail in this application.

[0161] In this embodiment, the memory 92 may include high-speed random access memory, and may also include non-volatile memory, such as at least one disk storage device or other volatile solid-state storage device. The processor 93 may be a central processing unit (CPU), an application-specific integrated circuit (ASIC), a digital signal processor (DSP), an application-specific integrated circuit (ASIC), an off-the-shelf programmable gate array (FPGA), or other programmable logic devices. This application does not limit the structure and model of the memory 92 and processor 93 described above, and they can be flexibly adjusted according to actual needs.

[0162] It should be understood that, Figure 9The structure of the computer device shown does not constitute a limitation on the computer device in the embodiments of this application. In practical applications, the computer device may include more than Figure 9 The computer device may include additional components or combinations of certain components. For example, if the computer device is a terminal, the terminal may also include at least one input component such as a touch sensing unit for sensing touch events on a touch display panel, a keyboard, a mouse, a camera, a microphone, etc.; at least one output component such as a monitor, a speaker, a vibration mechanism, a lamp, etc.; an antenna; a sensor module; a power supply module, etc., which may be determined according to the type and function of the terminal. If the computer device is a microscope in a microscopic imaging system, it may also include basic components for realizing the microscope function, such as the body, lenses / objectives, etc., which will not be listed here.

[0163] Finally, it should be noted that the various embodiments in this specification are described in a progressive or parallel manner. Each embodiment focuses on describing the differences from other embodiments, and the same or similar parts between the embodiments can be referred to mutually. Regarding the computer devices, apparatuses, and computer-readable storage media disclosed in the embodiments, since they correspond to the methods disclosed in the embodiments, the descriptions are relatively simple, and relevant parts can be referred to the method section.

[0164] Those skilled in the art will further recognize that the units and algorithm steps of the various examples described in conjunction with the embodiments disclosed herein can be implemented in electronic hardware, computer software, or a combination of both. To clearly illustrate the interchangeability of hardware and software, the components and steps of the various examples have been generally described in terms of functionality in the foregoing description. Whether these functions are implemented in hardware or software depends on the specific application and design preconditions of the technical solution. Those skilled in the art can use different methods to implement the described functions for each specific application, but such implementation should not be considered beyond the scope of this application.

[0165] The above description of the disclosed embodiments enables those skilled in the art to make or use this application. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the core ideas or scope of this application. Therefore, this application is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. An image processing method for a microscope under arbitrary phase modulation, characterized in that, The method includes: The first intensity image and phase optical transfer function of the sample are obtained under a microscopic imaging system; the first intensity image is acquired after a frame mask is configured in the microscopic imaging system. The first intensity image is preprocessed to obtain the second intensity image; An effective transfer function is obtained by using multiple preset phase values ​​and the phase optical transfer function; Using the effective transfer function, the second intensity image is deconvolved to obtain the target image of the sample, as well as the target phase shift generated by the amplitude mask introduced by the microscopic imaging system; The target phase shift is associated with the system identifier of the microscopic imaging system and stored, so as to use the target phase shift to reconstruct images of other intensities acquired by the microscopic imaging system; The step of using the effective transfer function to perform a deconvolution operation on the second intensity image to obtain the target phase shift generated by the amplitude mask configured in the microscopic imaging system includes: The effective transfer function is deconvolved with the second intensity image to obtain the deconvolution results corresponding to each of the plurality of preset phase values; the deconvolution results can characterize the image contrast of the corresponding reconstructed image. Compare multiple deconvolution results and select a preset phase value corresponding to the deconvolution result that represents the highest image contrast. The selected preset phase value is determined as the target phase shift generated by configuring the amplitude mask in the microscopic imaging system.

2. The method of claim 1, wherein, The step of preprocessing the first intensity image to obtain the second intensity image includes: The first intensity image is subjected to background removal processing to obtain a background-removed intensity image; The background-removed intensity image is subjected to background normalization processing to obtain a second intensity image.

3. The method of claim 1, wherein, During the process of deconvolution of the second intensity image using the effective transfer function, the incident light on the sample under the microscopic imaging system is amplitude modulated to reconstruct the phase information or refractive index information of the sample; the amplitude modulation can cause any phase shift of the target to be determined.

4. The method according to claim 1 or 2, characterized in that, The phase optical transfer function includes a first optical transfer function corresponding to a first point diffusion function and a second optical transfer function corresponding to a second point diffusion function; the first point diffusion function and the second point diffusion function are respectively used to represent the relationship between the conjugate function corresponding to the second intensity image and the scattering potential of the sample; The process of obtaining an effective transfer function by utilizing multiple preset phase differences and the phase optical transfer function includes: Multiple preset phase values ​​can be obtained; An effective transfer function is generated using the multiple preset phase values, the first optical transfer function, and the second optical transfer function.

5. The method according to claim 4, characterized in that, The step of performing a deconvolution operation on the second intensity image using the effective transfer function includes: Based on the transformation relationship between the sample scattering potential, the point spread function, and the conjugate function of the second intensity image, the expression of the second intensity image is adjusted; the adjusted expression of the second intensity image can represent the convolution relationship between the point spread function and the sample scattering potential. The second intensity image is deconvolved using the effective transfer function.

6. The method of claim 1 or 2, wherein, The method further includes: Obtain the intensity image of the sample to be processed acquired under the microscopic imaging system, and retrieve the target phase shift associated with the system identifier of the microscopic imaging system; Based on the target phase shift, the preprocessed intensity image to be processed is reconstructed to obtain the target image of the sample being tested.

7. An image processing apparatus for a microscope under arbitrary phase modulation, characterized by, The device includes: The data acquisition module is used to acquire the first intensity image and phase optical transfer function of the sample under the microscopic imaging system; the first intensity image is acquired after a frame mask is introduced into the microscopic imaging system. The preprocessing module is used to preprocess the first intensity image to obtain the second intensity image; An effective transfer function acquisition module is used to obtain an effective transfer function using multiple preset phase differences and the phase optical transfer function; The deconvolution operation module is used to perform a deconvolution operation on the second intensity image using the effective transfer function to obtain the target image of the sample and the target phase difference generated by the amplitude mask introduced by the microscopic imaging system. The data storage module is used to associate the target phase difference with the system identifier of the microscopic imaging system and store it, so as to use the target phase difference to reconstruct images of other intensities acquired by the microscopic imaging system. The deconvolution operation module includes: The deconvolution operation unit is used to perform a deconvolution operation on the effective transfer function and the second intensity image to obtain the deconvolution result corresponding to each of the plurality of preset phase values; the deconvolution result can characterize the image contrast of the corresponding reconstructed image. The comparison unit is used to compare multiple deconvolution results and select a preset phase value corresponding to the deconvolution result that represents the highest image contrast. The target phase shift determination unit is used to determine the selected preset phase value as the target phase shift generated by configuring the amplitude mask in the microscopic imaging system.

8. A computer device, comprising: The computer device includes: Communication module; At least one memory is used to store a program for implementing the image processing method for a microscope under arbitrary phase modulation as described in any one of claims 1-6; At least one processor is configured to load and execute the program stored in the memory to implement the image processing method for a microscope under arbitrary phase modulation as described in any one of claims 1-6.

9. A computer-readable storage medium, characterized in that, It stores a plurality of computer instructions, which are loaded and executed by a processor to implement the image processing method for a microscope under arbitrary phase modulation as described in any one of claims 1-6.

Images