Differential polarization confocal microscopic imaging system and method of multi-focus channel

Abstract

The invention discloses a differential polarization confocal microscopic imaging system and method of a multi-focus channel, and relates to the technical field of optical microscopy. Comprising a light source emission module, a dichroscope (6), a two-dimensional galvanometer module, a scanning module, a sample acquisition module, a detection module and an imaging terminal (17). According to the invention, differential polarization imaging and a multi-focus channel confocal technology are combined, so that the imaging quality of the confocal microscopic system is further improved on the premise of ensuring the confocal imaging speed; meanwhile, differential imaging is introduced, so that the imaging contrast ratio can be greatly improved, and the confocal microscopic system can obtain a better imaging effect in the application field of thick tissue microscopic imaging.

Description

Technical Field

[0001] The present invention relates to the field of optical microscopy technology, and in particular to a differential polarization confocal microscopy imaging system and method for multi-focus channels. Background Art

[0002] Confocal microscopy is a high-resolution imaging technology widely used in biology, materials science and medical research. Compared with traditional microscopes, confocal microscopy can obtain higher image clarity and contrast through point scanning and optical sectioning technology. This microscope uses a point light source to illuminate the sample and filters the scattered light of the non-focal plane through a pinhole, thereby realizing three-dimensional reconstruction and multi-level imaging of the sample. In addition, differential polarization technology can effectively reduce these interferences, thereby further improving the image contrast and detail recognition ability of the confocal microscope. Therefore, the differential polarization technology of multi-focus channels is applied to confocal microscopy, which helps to achieve high-resolution and high-contrast imaging in the fields of biomedicine, materials science, etc. Background technology includes the application of laser scanning technology, fluorescent dye labeling and highly sensitive photodetectors, which further improves the imaging quality and analysis accuracy. The development of these technologies makes confocal microscopy an indispensable tool in modern scientific research.

[0003] Traditional confocal microscope systems have the following problems when imaging: (1) Background noise interference. Traditional systems are easily affected by background scattered light, resulting in reduced image quality and difficulty in clearly distinguishing fine structures in the sample. (2) Insufficient contrast. In complex samples, the lack of effective contrast may make it difficult to distinguish the target object from the background. (3) Imaging speed limitation. Traditional confocal microscopes often require point-by-point scanning, and the imaging speed is relatively slow, which limits the ability of dynamic observation. (4) Photodamage and photobleaching. Under long-term exposure, the sample may be damaged by light, resulting in photobleaching, which affects the observation results. (5) Limited depth resolution. For thick samples, traditional confocal microscopes will face the problem of reduced resolution when imaging in deep layers.

[0004] Therefore, a differential polarization confocal microscopy imaging system and method with multiple focal channels is proposed. By introducing multiple focal channels and differential polarization technology, the above problems can be effectively overcome, which is an issue that technical personnel in this field urgently need to solve. Summary of the invention

[0005] In view of this, the present invention provides a differential polarization confocal microscopy imaging system and method with multiple focal channels, which combines differential polarization imaging with the confocal technology of multiple focal channels, thereby further improving the imaging quality of the confocal microscopy system while ensuring the confocal imaging speed.

[0006] In order to achieve the above object, the present invention adopts the following technical solution:

[0007] A differential polarization confocal microscopic imaging system with multiple focal channels, comprising:

[0008] Light source emission module, dichroic mirror, two-dimensional galvanometer module, scanning module, sample collection module, detection module and imaging terminal;

[0009] A light source transmitting module is used to transmit light beams of multiple polarization states to the two-dimensional galvanometer module, and a dichroic mirror is arranged on the light path between the light source transmitting module and the two-dimensional galvanometer module;

[0010] A two-dimensional galvanometer module is used to scan light beams in multiple polarization states to obtain light beams scanned by the galvanometer;

[0011] A scanning module, used for focusing the light beam scanned by the galvanometer onto the sample collection module;

[0012] The sample collection module is used for multi-point focusing on the microscopic imaging sample, that is, exciting multi-point fluorescence signals to obtain multi-focal fluorescence signals;

[0013] The multi-focal fluorescence signal returns along the original path, passes through the scanning module and the two-dimensional galvanometer module in sequence, and finally reaches the surface of the dichroic mirror. The multi-focal fluorescence signal is reflected on the surface of the dichroic mirror to obtain the reflected fluorescence signal;

[0014] A detection module, used for receiving the polarized fluorescence signal, and sending the polarized fluorescence signal to the imaging terminal after processing;

[0015] The imaging terminal is used to perform sample imaging on the processed polarized fluorescence signal to obtain a sample image.

[0016] In the above system, optionally, the light source emission module includes a laser, and a beam splitter positioning plate, an excitation end polarizer, an excitation end correction lens and an excitation end filter arranged along the laser emission direction of the laser;

[0017] Laser, used for emitting laser light;

[0018] A beam splitting positioning plate, used for splitting the laser light emitted by the laser into multiple beams to form a multi-beam channel;

[0019] An excitation end polarizer is used to modulate the polarization states of the multiple light beams to generate light beams of multiple polarization states;

[0020] The excitation end correction lens is used to calibrate the beams of multiple polarization states in multiple channels and couple them to the two-dimensional galvanometer module to achieve joint scanning of multiple channels;

[0021] Excitation end filter, used to eliminate the interference of stray light other than the excitation light wavelength;

[0022] In the above system, optionally, the two-dimensional galvanometer module includes an X-axis galvanometer and a Y-axis galvanometer;

[0023] The X-axis galvanometer is responsible for one-dimensional scanning in the x-axis direction, and the Y-axis galvanometer is responsible for one-dimensional scanning in the y-axis direction. The one-dimensional scanning in the x-axis direction and the one-dimensional scanning in the y-axis direction are combined to form a two-dimensional scan. Light beams in multiple polarization states pass through the two-dimensional galvanometer module to realize the scanning process.

[0024] In the above system, optionally, the scanning module includes a scanning lens and a correction tube lens arranged along the optical path;

[0025] A scanning lens is used to initially focus the multi-channel light beam scanned by the two-dimensional galvanometer module;

[0026] The correction tube lens is used to cooperate with the microscope objective lens to reduce the focused light spot of the scanning lens and irradiate the light spot to the microscopic imaging sample.

[0027] In the above system, optionally, the sample collection module includes a microscope objective lens and a microscopic imaging sample arranged along the optical path;

[0028] The microscope objective lens is used to cooperate with the correction tube lens to reduce the initial focusing spot of the scanning lens, and then irradiate the microscopic imaging sample.

[0029] In the above system, optionally, the detection module includes a receiving end filter, a receiving end correction lens, a polarization detector and a light intensity detector array arranged along the reflection light path of the dichroic mirror;

[0030] Since the wavelength of the fluorescence signal is greater than the wavelength of the excitation light, the fluorescence signal is reflected on the surface of the dichroic mirror. The reflected fluorescence signal passes through the receiving end filter and the receiving end correction lens, and then the polarization state is modulated by the polarization detector, and the light intensity detector array extracts the signal.

[0031] A differential polarization confocal microscopy imaging method for multiple focal channels, applied to a differential polarization confocal microscopy imaging system for multiple focal channels as described in any one of the above, comprises the following contents:

[0032] After the laser is split, it is fixed to the splitter positioning plate to obtain multiple beams to form a multi-beam channel;

[0033] The multiple light beams are polarized by a polarizer at the excitation end to generate light beams with multiple polarization states;

[0034] The light beams in multiple polarization states then pass through the excitation end correction lens, the excitation end filter, and the dichroic mirror in sequence before reaching the two-dimensional galvanometer module, where the X-axis galvanometer is responsible for one-dimensional scanning in the x-axis direction, and the Y-axis galvanometer is responsible for one-dimensional scanning in the y-axis direction. The one-dimensional scanning in the x-axis direction and the one-dimensional scanning in the y-axis direction are combined to form a two-dimensional scan to realize the scanning process;

[0035] The light beam scanned by the two-dimensional galvanometer module is then passed through the scanning lens, correction tube lens, and microscope objective lens to achieve multi-point focusing on the microscopic imaging sample, that is, to stimulate multi-focal fluorescence signals;

[0036] The multi-focus fluorescence signal returns along the original path, passes through the microscope objective, correction tube lens, scanning lens, Y-axis galvanometer, X-axis galvanometer, and finally reaches the surface of the dichroic mirror;

[0037] The multi-focal fluorescence signals produce a reflection effect on the surface of the dichroic mirror;

[0038] The reflected fluorescence signal passes through the receiving end filter and the receiving end correction lens, and then the polarization state is modulated by the polarization detector. Finally, the signal is extracted by the light intensity detector array.

[0039] The extracted signals are fused through the imaging terminal to obtain the microscopic imaging sample image.

[0040] In the above method, optionally, during the imaging process, the excitation end polarizer and the polarization detector will be combined with each other multiple times, and multiple groups of polarization imaging results will be obtained through multiple adjustments. Finally, the multiple groups of results will be differentially processed to obtain the final imaging result.

[0041] It can be seen from the above technical solutions that, compared with the prior art, the present invention provides a multi-focal channel differential polarization confocal microscopy imaging system and method, which has the following beneficial effects:

[0042] By combining differential polarization imaging and confocal technology with multiple focal channels, the imaging quality of the confocal microscopy system can be further improved while ensuring the confocal imaging speed. At the same time, the introduction of differential imaging can greatly improve the imaging contrast, enabling the confocal microscopy system to achieve better imaging effects in the application field of thick tissue microscopy. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0044] Figure 1 An optical path diagram of a differential polarization confocal microscopic imaging system with multiple focal channels disclosed in the present invention;

[0045] Figure 2 A differential polarization confocal microscopy imaging flow chart of a multi-focus channel disclosed in the present invention;

[0046] Figure 3 It is a control structure framework diagram of the differential polarization confocal microscopic imaging system with multiple focal channels disclosed in the present invention;

[0047] Figure 4 A schematic diagram of the differential polarization fusion effect disclosed in an embodiment of the present invention;

[0048] Figure 5 It is a schematic diagram of multi-focus channel field of view image fusion disclosed in an embodiment of the present invention. DETAILED DESCRIPTION

[0049] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0050] In this application, relational terms such as first and second, etc. are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. The terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of more restrictions, the elements defined by the sentence "including one..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0051] Reference Figure 1 As shown, the present invention discloses a differential polarization confocal microscopic imaging system with multiple focal channels, comprising:

[0052] Light source emission module, dichroic mirror 6, two-dimensional galvanometer module, scanning module, sample collection module, detection module and imaging terminal 17;

[0053] A light source transmitting module is used to transmit light beams of multiple polarization states to the two-dimensional galvanometer module, and a dichroic mirror 6 is arranged on the light path between the light source transmitting module and the two-dimensional galvanometer module;

[0054] A two-dimensional galvanometer module is used to scan light beams in multiple polarization states to obtain light beams scanned by the galvanometer;

[0055] A scanning module, used for focusing the light beam scanned by the galvanometer onto the sample collection module;

[0056] A sample collection module, used for multi-point focusing on the microscopic imaging sample 12, that is, exciting multi-point fluorescence signals to obtain multi-focal fluorescence signals;

[0057] The multi-focus fluorescence signal returns along the original path, passes through the scanning module and the two-dimensional galvanometer module in sequence, and finally reaches the surface of the dichroic mirror 6. The multi-focus fluorescence signal is reflected on the surface of the dichroic mirror 6 to obtain the reflected fluorescence signal;

[0058] A detection module, used for receiving the polarized fluorescence signal, and sending the polarized fluorescence signal to the imaging terminal 17 after processing;

[0059] The imaging terminal 17 is used to perform sample imaging on the processed polarized fluorescence signal to obtain a sample image.

[0060] Furthermore, the light source emission module includes a laser 1, and a light splitting positioning plate 2, an excitation end polarizer 3, an excitation end correction lens 4 and an excitation end filter 5 arranged along the laser emission direction of the laser 1;

[0061] A laser 1, used for emitting laser light;

[0062] A beam splitting positioning plate 2, used for splitting the laser light emitted by the laser 1 into multiple beams to form a multi-beam channel;

[0063] The excitation end polarizer 3 is used to modulate the polarization states of the multiple light beams to generate light beams of multiple polarization states;

[0064] The excitation end correction lens 4 is used to perform beam calibration of multiple channels on beams of multiple polarization states and is coupled to a two-dimensional galvanometer module (between the X-axis galvanometer 7 and the Y-axis galvanometer 8) to achieve joint scanning of multiple channels;

[0065] The excitation end filter 5 is used to eliminate the interference of stray light other than the excitation light wavelength.

[0066] Further, the two-dimensional galvanometer module includes an X-axis galvanometer 7 and a Y-axis galvanometer 8;

[0067] The X-axis galvanometer 7 is responsible for one-dimensional scanning in the x-axis direction, and the Y-axis galvanometer 8 is responsible for one-dimensional scanning in the y-axis direction. The one-dimensional scanning in the x-axis direction and the one-dimensional scanning in the y-axis direction are combined to form a two-dimensional scanning. Light beams in multiple polarization states pass through the two-dimensional galvanometer module to realize the scanning process.

[0068] Further, the scanning module includes a scanning lens 9 and a correction tube lens 10 arranged along the optical path;

[0069] Scanning lens 9, used to initially focus the multi-channel light beam scanned by the two-dimensional galvanometer module;

[0070] The correction tube lens 10 is used to cooperate with the infinitely conjugated microscope objective lens 11 to reduce the focused light spot of the scanning lens 9 and irradiate the light spot to the microscopic imaging sample 12.

[0071] Further, the sample collection module includes a microscope objective lens 11 and a microscopic imaging sample 12 arranged along the optical path;

[0072] The microscope objective lens 11 is used to cooperate with the correction tube lens 10 to reduce the initial focusing spot of the scanning lens 9 and then irradiate the microscope imaging sample 12.

[0073] Further, the detection module includes a receiving end filter 13, a receiving end correction lens 14, a polarization detector 15 and a light intensity detector array 16 arranged along the reflection light path of the dichroic mirror 6;

[0074] There are multiple light intensity detector arrays 16, which can be set as needed.

[0075] Since the wavelength of the fluorescence signal is greater than the wavelength of the excitation light, the fluorescence signal is reflected on the surface of the dichroic mirror 6. The reflected fluorescence signal passes through the receiving end filter 13 and the receiving end correction lens 14, and then the polarization state is modulated by the polarization detector 15, and the light intensity detector array 16 extracts the signal.

[0076] The principle of the differential polarization confocal microscopic imaging system with multiple focus channels of the present invention is as follows: a laser 1 obtains multiple light beams after passing through a splitting positioning plate 2, and the multiple light beams are polarized by an excitation end polarizer 3, and then sequentially pass through an excitation end correction lens 4, an excitation end filter 5 and a dichroic mirror 6 to reach a two-dimensional galvanometer module (composed of an X-axis galvanometer 7 and a Y-axis galvanometer 8). The light beam passes through the two-dimensional galvanometer module to realize a scanning process. The light beam after passing through the two-dimensional galvanometer module can realize multi-point focusing on a microscopic imaging sample 12 by passing through a scanning lens 9, a correction tube lens 10 and a microscope objective lens 11, that is, it can excite multi-point fluorescence signals. The multi-focus fluorescence signal returns to the surface of the dichroic mirror 6 along the original path. Since the fluorescence signal has a longer wavelength than the excitation light, it has a reflection effect on the surface of the dichroic mirror 6. The reflected fluorescence signal passes through a receiving end filter 13 and a receiving end calibration lens 14, and then is polarized by a polarization detector 15. Finally, the light intensity detection array 16 extracts the signal, and the extracted signal can generate a sample image through the imaging terminal 17. During the imaging process, the polarizer and polarization detector are adjusted multiple times to obtain multiple sets of polarization imaging results, and finally the multiple sets of results are differentially processed to obtain the best imaging result.

[0077] During the imaging process, the excitation end polarizer 3 and the polarization detector 15 will be combined with each other for multiple times, and multiple groups of polarization imaging results will be obtained in multiple adjustments. Finally, the multiple groups of results will be differentially processed to obtain the best thick tissue imaging result.

[0078] See also Figure 2 As shown, the present invention also discloses a differential polarization confocal microscopy imaging method of multiple focal channels, which is applied to the differential polarization confocal microscopy imaging system of multiple focal channels described above, and includes the following contents:

[0079] After the laser 1 is split, it is fixed to the splitting positioning plate 2 to obtain multiple beams to form a multi-beam channel;

[0080] The multiple light beams are polarized by the excitation end polarizer 3 to generate light beams with multiple polarization states;

[0081] The light beams in multiple polarization states then pass through the excitation end correction lens 4, the excitation end filter 5, and the dichroic mirror 6 in sequence before reaching the two-dimensional galvanometer module, wherein the X-axis galvanometer is responsible for one-dimensional scanning in the x-axis direction, and the Y-axis galvanometer is responsible for one-dimensional scanning in the y-axis direction. The one-dimensional scanning in the x-axis direction and the one-dimensional scanning in the y-axis direction are combined to form a two-dimensional scan to realize the scanning process;

[0082] The light beam scanned by the two-dimensional galvanometer module is then passed through the scanning lens 9, the correction tube lens 10, and the microscope objective lens 11 to achieve multi-point focusing on the microscopic imaging sample 12, that is, to stimulate multi-focal fluorescence signals;

[0083] The multi-focus fluorescence signal returns along the original path, passes through the microscope objective lens 11, the correction tube lens 10, the scanning lens 9, the Y-axis galvanometer 8, the X-axis galvanometer 7, and finally reaches the surface of the dichroic mirror 6;

[0084] The multi-focal fluorescence signals produce a reflection effect on the surface of the dichroic mirror 6;

[0085] The reflected fluorescence signal passes through the receiving end filter 13 and the receiving end correction lens 14, and then the polarization state is modulated by the polarization detector 15, and finally, the signal is extracted by the light intensity detector array 16;

[0086] The extracted signals are fused through the imaging terminal 17 to obtain an image of the microscopic imaging sample 12 .

[0087] Furthermore, during the imaging process, the excitation end polarizer 3 and the polarization detector 15 will be combined with each other multiple times, and multiple groups of polarization imaging results will be obtained through multiple adjustments. Finally, the multiple groups of results will be differentially processed to obtain the final imaging result.

[0088] See also Figure 3 As shown, the present invention also discloses a control structure framework diagram of a differential polarization confocal microscopic imaging system with multiple focus channels, including: a microscope processing system, and a laser light intensity control module, a PMT array acquisition module, a transmitting and receiving glass slide rotation control module, and an electric galvanometer motor control module connected to the microscope processing system;

[0089] Microscope processing system, used to process the collected light intensity signals into microscopic imaging pictures, the processing process includes but is not limited to focus control, image reconstruction, multi-channel image stitching and polarization difference image processing;

[0090] A laser light intensity control module, used to control the intensity of the laser emitted by the laser 1;

[0091] A PMT array acquisition module, used to control the light intensity detector array 16 to receive and extract the fluorescence signal;

[0092] The transmitting and receiving slide rotation control module is used to switch the polarization state of the excitation light and the polarization state of the emitted fluorescence. The multiple states are switched to obtain fluorescence information under different polarization states, which is then used for subsequent polarization differential processing in the microscope processing system to obtain images with higher contrast.

[0093] The electric galvanometer motor control module is used to control the scanning of the two-dimensional galvanometer module (X-axis galvanometer 7 and Y-axis galvanometer 8) to achieve scanning imaging of multi-channel light beams on the sample plane.

[0094] In a specific embodiment, see Figure 4 The figure shows a schematic diagram of the differential polarization fusion effect. The fluorescent microtube image collected at 0° polarization state and the same sample fluorescent microtube image collected at 90° polarization state are subjected to differential polarization fusion processing to obtain a polarization fused image, and the image quality is higher than that of the image without differential polarization fusion.

[0095] The reference process of polarization difference fusion processing is as follows: data acquisition, collecting image data at different polarization angles (such as 0, 45, 90, 135), and recording the light intensity at different pixel positions at different polarization angles; polarization difference map calculation, performing differential calculation on images under different polarization states, obtaining the difference in polarization states in the sample and performing contrast enhancement to improve the contrast of tiny structures in the sample; denoising and filtering, applying denoising filters, such as Gaussian filtering, median filtering, edge enhancement filtering, etc., to process noise and obtain the reconstructed image; finally, quantitative analysis and visualization, performing quantitative analysis on the reconstructed image.

[0096] In a specific embodiment, see Figure 5 Shown is a schematic diagram of multi-focus channel field of view image fusion, the fluorescence images of 6 sub-field of view channels and the final fused full-field grayscale image and full-field fluorescence image.

[0097] Each embodiment in this specification is described in a progressive manner, and the same or similar parts between the embodiments can refer to each other, and each embodiment focuses on the differences from other embodiments. In particular, for the system or system embodiment, since it is basically similar to the method embodiment, the description is relatively simple, and the relevant parts can refer to the partial description of the method embodiment. The system and system embodiments described above are merely schematic, wherein the units described as separate components may or may not be physically separated, and the components displayed as units may or may not be physical units, that is, they may be located in one place, or they may be distributed on multiple network units. Some or all of the modules may be selected according to actual needs to achieve the purpose of the scheme of this embodiment. Ordinary technicians in this field can understand and implement it without creative work.

[0098] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A differential polarization confocal microscopy imaging system with multiple focal channels, characterized in that: include: A light source emission module, a dichroic mirror (6), a two-dimensional galvanometer module, a scanning module, a sample collection module, a detection module and an imaging terminal (17); A light source transmitting module is used to transmit light beams of multiple polarization states to the two-dimensional galvanometer module, and a dichroic mirror (6) is arranged on the light path between the light source transmitting module and the two-dimensional galvanometer module; A two-dimensional galvanometer module is used to scan light beams in multiple polarization states to obtain light beams scanned by the galvanometer; A scanning module, used for focusing the light beam scanned by the galvanometer onto the sample collection module; A sample collection module, used for multi-point focusing on a microscopic imaging sample (12), that is, exciting multi-point fluorescence signals to obtain multi-focal fluorescence signals; The multi-focal fluorescence signal returns along the original path, passes through the scanning module and the two-dimensional galvanometer module in sequence, and finally reaches the surface of the dichroic mirror (6). The multi-focal fluorescence signal is reflected on the surface of the dichroic mirror (6) to obtain a reflected fluorescence signal; A detection module, used for receiving the polarized fluorescence signal, and processing the polarized fluorescence signal and sending it to the imaging terminal (17); The imaging terminal (17) is used to perform sample imaging on the processed polarized fluorescence signal to obtain a sample image.

2. The differential polarization confocal microscopy imaging system with multiple focal channels according to claim 1, characterized in that: The light source emission module comprises a laser (1), and a light splitting positioning plate (2), an excitation end polarizer (3), an excitation end correction lens (4) and an excitation end filter (5) arranged along the laser emission direction of the laser (1); A laser (1), used for emitting laser light; A beam splitting positioning plate (2) is used to split the laser light emitted by the laser (1) into multiple beams to form a multi-beam channel; An excitation end polarizer (3) is used to modulate the polarization states of the multiple light beams to generate light beams with multiple polarization states; An excitation end correction lens (4) is used to perform beam calibration of multiple channels on beams of multiple polarization states and couple them to a two-dimensional galvanometer module to achieve joint scanning of multiple channels; The excitation end filter (5) is used to eliminate the interference of stray light other than the excitation light wavelength.

3. The differential polarization confocal microscopy imaging system with multiple focal channels according to claim 2, characterized in that: The two-dimensional galvanometer module comprises an X-axis galvanometer (7) and a Y-axis galvanometer (8); The X-axis galvanometer (7) is responsible for one-dimensional scanning in the x-axis direction, and the Y-axis galvanometer (8) is responsible for one-dimensional scanning in the y-axis direction. The one-dimensional scanning in the x-axis direction and the one-dimensional scanning in the y-axis direction are combined to form a two-dimensional scanning. Light beams in multiple polarization states pass through the two-dimensional galvanometer module to realize the scanning process.

4. The differential polarization confocal microscopy imaging system with multiple focal channels according to claim 3, characterized in that: The scanning module comprises a scanning lens (9) and a correction tube lens (10) arranged along the optical path; A scanning lens (9) is used to initially focus the multi-channel light beam scanned by the two-dimensional galvanometer module; The correction tube lens (10) is used to cooperate with the microscope objective lens (11) to reduce the focused light spot of the scanning lens (9) so that the light spot is irradiated onto the microscopic imaging sample (12).

5. The differential polarization confocal microscopy imaging system with multiple focal channels according to claim 4, characterized in that: The sample collection module comprises a microscope objective lens (11) and a microscopic imaging sample (12) arranged along the optical path; The microscope objective lens (11) is used to cooperate with the correction tube lens (10) to reduce the initial focus spot of the scanning lens (9) and then irradiate the microscope imaging sample (12).

6. The differential polarization confocal microscopy imaging system with multiple focal channels according to claim 5, characterized in that: The detection module comprises a receiving end filter (13), a receiving end correction lens (14), a polarization detector (15) and a light intensity detector array (16) arranged along the reflection light path of the dichroic mirror (6); Since the wavelength of the fluorescence signal is greater than the wavelength of the excitation light, the fluorescence signal is reflected on the surface of the dichroic mirror (6). The reflected fluorescence signal passes through a receiving end filter (13) and a receiving end correction lens (14), and then is polarized by a polarization detector (15) for modulation of the polarization state, and a light intensity detector array (16) is used for signal extraction.

7. A differential polarization confocal microscopy imaging method with multiple focal channels, characterized in that: A differential polarization confocal microscopy imaging system with multiple focal channels applied to any one of claims 1 to 6, comprising the following contents: The laser light emitted by the laser (1) is fixed to the light splitting positioning plate (2) after light splitting, and a plurality of light beams are obtained to form a multi-beam channel; The multiple light beams are subjected to polarization state modulation through an excitation end polarizer (3) in order to generate light beams with multiple polarization states; The light beams in multiple polarization states then pass through the excitation end correction lens (4), the excitation end filter (5), and the dichroic mirror (6) in sequence before reaching the two-dimensional galvanometer module, wherein the X-axis galvanometer is responsible for one-dimensional scanning in the x-axis direction, and the Y-axis galvanometer is responsible for one-dimensional scanning in the y-axis direction. The one-dimensional scanning in the x-axis direction and the one-dimensional scanning in the y-axis direction are combined to form a two-dimensional scan, thereby realizing the scanning process; The light beam scanned by the two-dimensional galvanometer module is then passed through a scanning lens (9), a correction tube lens (10), and a microscope objective lens (11) to achieve multi-point focusing on a microscopic imaging sample (12), that is, to excite multi-focal fluorescence signals; The multi-focus fluorescence signal returns along the original path, passes through the microscope objective lens (11), the correction tube lens (10), the scanning lens (9), the Y-axis galvanometer (8), the X-axis galvanometer (7), and finally reaches the surface of the dichroic mirror (6); The multi-focal fluorescence signals are reflected on the surface of the dichroic mirror (6); The reflected fluorescence signal passes through a receiving end filter (13) and a receiving end correction lens (14), and then is modulated in polarization state by a polarization detector (15). Finally, the signal is extracted by a light intensity detector array (16); The extracted signals are subjected to image fusion via an imaging terminal (17) to obtain an image of the microscopic imaging sample (12).

8. The differential polarization confocal microscopy imaging method with multiple focal channels according to claim 7, characterized in that: During the imaging process, the excitation end polarizer (3) and the polarization detector (15) are combined with each other multiple times, and multiple groups of polarization imaging results are obtained by performing multiple adjustments. Finally, the multiple groups of results are differentially processed to obtain the final imaging result.

Images