Preparation method of small sample pathological digital slide

Abstract

The invention relates to a preparation method of a pathological digital slide for a tiny sample, which realizes quick positioning and focusing by comparing a focusing mode and collecting a digital image of the tiny sample, and solves the problems of positioning and focusing of the tiny sample under a slide scanner. The invention provides a more effective tool for pathologists and medical researchers to research pathological characteristics of small samples. The micro samples can be analyzed and studied more accurately, the occurrence mechanism, the development process, the response to treatment and the like of diseases can be deeply known, powerful support is provided for medical research, and continuous development of the pathology department is promoted. The small samples can be better processed, doctors can be helped to more accurately judge the properties and stages of tumors, and a basis is provided for formulating a treatment scheme. The prepared high-quality pathological digital slide image is easier to transmit and share, and remote consultation and academic communication between pathologists are facilitated.

Description

Technical Field

[0001] The present disclosure relates to the field of biological experiment technology, and in particular to a method for preparing a micro-sample pathology digital slide. Background Art

[0002] Pathology digital slides (WSI) are a technology product that converts traditional pathology slides into digital images through digital scanning and processing. Pathology digital slides are widely used in the current medical, scientific research and teaching fields. In terms of storage and management, traditional pathology slides take up a lot of physical space and are easily damaged, faded or lost after long-term storage. Digital slides are stored in computer hard drives, servers or cloud storage in digital form. This not only greatly saves physical storage space, but also makes it easy to achieve orderly management of massive pathology images. Through the database system, the required pathology slice images can be quickly retrieved, and the stability of digital storage ensures that the image quality is not affected by time and can be called at any time.

[0003] Digital slides are an important aid to improving diagnostic accuracy. Computer software can perform detailed image analysis and processing, such as quantitative analysis of cell morphology, size, nuclear-cytoplasmic ratio, tissue structure, staining characteristics, etc. This quantitative data provides a more objective and accurate basis for pathologists' diagnosis. At the same time, digital slides facilitate multi-slice comparative observation, can clearly present the manifestations of lesions at different levels, and can also achieve three-dimensional reconstruction, showing the pathological structure from a three-dimensional perspective, allowing doctors to have a more comprehensive understanding of the lesions and reduce misdiagnosis and missed diagnosis.

[0004] Information sharing and communication are another outstanding advantage of digital slides. With the help of the Internet, digital slides can be quickly transmitted between pathologists in different hospitals and regions. In remote consultations, experts can intuitively view pathological sections without touching the slides, and make diagnoses and discussions as if they were on site. In academic exchange activities, researchers can easily share special or typical cases to promote the dissemination and exchange of medical knowledge.

[0005] Digital slides have laid the foundation for the application of artificial intelligence in the field of pathological diagnosis. A large amount of digital slide data has become a valuable material for training artificial intelligence algorithms. By learning the patterns of different pathological characteristics, the algorithms can provide references for doctors during the diagnosis process, especially for diagnostic work with large workload and high repetitiveness, which can effectively improve the efficiency and quality of diagnosis.

[0006] In the field of medical education, digital slides have shown good teaching effects. Teachers can use them to show students clear details of pathological sections, and through the annotation function of the software, clearly point out key pathological features to guide students to learn. At the same time, it is more convenient to use digital slides for examinations and assessments. Teachers can quickly prepare assessment content and accurately assess students' mastery of pathological knowledge, thereby improving teaching quality.

[0007] At present, the main problem in the research of ultra-small pathological samples such as organoids, clinical punctures, and micro-organs is their small size, which is usually in the range of several hundred microns to several millimeters. The resolution of digital slide images produced by slide scanners can reach the micron level. However, before starting the scanning work, the slide scanner cannot accurately identify tiny tissue samples with a diameter of less than 0.5mm. Such small pathological samples pose a severe challenge to the preparation of pathological digital slides. If the sample is too small (less than 0.5mm in diameter), it is difficult to locate and cannot provide sufficient focus (≥5), resulting in blur and loss of focus, scanning blind spots, and serious background interference, which in turn leads to the failure of pathological digital slide preparation (such as attached Figure 1 and attached Figure 2 The digital slide image of the 0.5 mm liver sample shown in the figure). When the liver sample from the same animal is 5 mm in diameter, a clear HE-stained digital slide image with no background interference can be obtained (see the attached Figure 3 and attached Figure 4 Digital slide image of a 5 mm liver sample shown).

[0008] Therefore, tiny samples are difficult to locate and focus on a slide scanner, and it is necessary to improve the preparation method of digital slides for pathology of tiny samples, so as to solve the problem that tiny samples are difficult to locate and focus on a slide scanner. By improving the slide preparation method, the samples can be easier to identify and align on the slide. Summary of the invention

[0009] In order to solve the above problems, the present application proposes a method for preparing a micro-sample pathology digital slide, which locates and displays the micro-sample by adding a contrast enhancer to the micro-sample or slicing it together with a large sample.

[0010] On the one hand, the present application provides a method for preparing a micro-sample pathology digital slide, comprising the following steps:

[0011] S1. Extract and pre-process tiny samples from animal model tissues;

[0012] S2, preparing a sample positioning marker and slicing it together with the micro-sample to form a pathology digital slide of the micro-sample;

[0013] S3, digitally scanning the pathology digital slide, positioning and focusing the micro-sample based on the sample positioning marker, and acquiring a WSI slice image of the micro-sample;

[0014] S4. Perform image processing on the WSI slice image, and locate the tiny sample based on the sample positioning marker.

[0015] On the other hand, the present application provides a method for preparing a micro-sample pathology digital slide, comprising the following steps:

[0016] S1. Extract and pre-process tiny samples from animal model tissues;

[0017] S2, slicing the tiny sample to form a pathology digital slide of the tiny sample, and adding a sample positioning marker on the pathology digital slide to provide a position marker for positioning and focusing the tiny sample on the pathology digital slide;

[0018] S3, digitally scanning the pathology digital slide, positioning and focusing the micro-sample based on the sample positioning marker, and acquiring a WSI slice image of the micro-sample;

[0019] S4. Perform image processing on the WSI slice image, and locate the tiny sample based on the sample positioning marker.

[0020] Technical effects of the present invention:

[0021] This application uses a contrast focusing method to collect digital images of tiny samples, achieve rapid positioning and focusing, solve the positioning and focusing problems of tiny samples under a slide scanner, reduce scanning blind areas, and improve image quality, providing strong support for pathological analysis of tiny samples.

[0022] The present invention has the following technical advantages:

[0023] 1. Technological improvements and breakthroughs:

[0024] Improve positioning and focusing accuracy: In the preparation and observation of conventional pathology slides, tiny samples are difficult to accurately position and focus on under slide scanners due to their extremely small size. This inventive method improves the preparation process, such as using large samples to assist in positioning and adding special marks, so that tiny samples can be clearly marked on the slide, so that the sample position can be quickly and accurately found and focused during the scanning process, greatly improving the convenience and accuracy of sample observation and reducing the risk of misdiagnosis or missed diagnosis due to positioning difficulties.

[0025] Reduce scanning blind areas: Tiny samples are easily unevenly distributed on the slide, causing some areas to be missed during scanning, forming scanning blind areas. New preparation methods can make the samples more evenly distributed on the slide, or through special slide preparation technology to ensure that the scanner can fully obtain the sample information, avoiding the impact of missing information on diagnosis and research results.

[0026] Enhanced image quality: The image information of tiny samples is limited, and the contrast with the background may not be high. By adding appropriate contrast-enhancing substances during the preparation process, such as adding brightly stained organs such as the pancreas, kidneys, and liver, or optimizing the sample processing steps, enhancing the sample color concentration, and extending the staining time, the sample staining contrast is more obvious, which can highlight the pathological characteristics of the sample, make the distinction between the sample and the background more obvious, and improve the clarity and recognition of the image, allowing pathologists to more accurately observe and analyze the pathological characteristics of the sample.

[0027] 2. Scientific research and medical applications:

[0028] Promoting the development of pathological research: It provides a more effective tool for pathologists and medical researchers to study the pathological characteristics of tiny samples. In medical research, only very small samples can be obtained in many cases, such as fine needle aspiration samples, micro-tissue sections, etc. This invention method enables these tiny samples to be analyzed and studied more accurately, which helps to gain a deeper understanding of the occurrence mechanism, development process and response to treatment of the disease, etc., providing strong support for medical research and promoting the continuous development of pathology.

[0029] Assisted disease diagnosis: In clinical diagnosis, for some diseases where it is difficult to obtain a large number of samples, the micro-sample pathology digital slide preparation method can improve the accuracy and reliability of diagnosis. For example, in the early diagnosis of tumors, sometimes only a small amount of cell samples can be obtained through fine needle puncture. This method can better process these tiny samples, help doctors more accurately determine the nature and stage of the tumor, and provide a basis for formulating treatment plans.

[0030] Facilitates remote consultation and data sharing: The high-quality digital pathology slide images prepared are easier to transmit and share, which facilitates remote consultation and academic exchanges between pathologists. Doctors in different regions can share image information of tiny sample pathology digital slides through the network, discuss cases together, and improve the level and efficiency of diagnosis. At the same time, it is also conducive to the establishment of a pathology database, providing basic data for large-scale clinical research and disease monitoring.

[0031] Further features and aspects of the present disclosure will become apparent from the following detailed description of exemplary embodiments with reference to the attached drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments, features, and aspects of the disclosure and, together with the description, serve to explain the principles of the disclosure.

[0033] Figure 1 Schematic diagram of a digital slide image of a 0.5 mm liver sample;

[0034] Figure 2 Shown as Figure 1 An enlarged schematic diagram of

[0035] Figure 3 A schematic diagram of a digital slide image of a 5 mm liver sample is shown;

[0036] Figure 4 Shown as Figure 3 An enlarged schematic diagram of

[0037] Figure 5 Shown is a schematic diagram of imaging of a large sample (lower part) and a small sample (upper part) of the liver in a low-power image of the WS I of the present invention;

[0038] Figure 6 Shown as Figure 5 Schematic diagram of WS I magnification of small and medium samples;

[0039] Figure 7 Shown as Figure 5 Schematic diagram of WS I magnification of medium and large samples;

[0040] Figure 8 The figure shows a low-magnification digital slide of the optic nerve of an I CR mouse showing pathological tissue of the optic nerve;

[0041] Fig. 9 A high-magnification digital slide of the optic nerve of an I CR mouse is shown, showing a schematic diagram of the pathological tissue of the optic nerve. DETAILED DESCRIPTION

[0042] Various exemplary embodiments, features and aspects of the present disclosure will be described in detail below with reference to the accompanying drawings. The same reference numerals in the accompanying drawings represent elements with the same or similar functions. Although various aspects of the embodiments are shown in the accompanying drawings, the drawings are not necessarily drawn to scale unless otherwise specified.

[0043] The word “exemplary” is used exclusively herein to mean “serving as an example, example, or illustration.” Any embodiment described herein as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments.

[0044] In addition, in order to better illustrate the present disclosure, numerous specific details are given in the following specific embodiments. It should be understood by those skilled in the art that the present disclosure can also be implemented without certain specific details. In some examples, means, components and circuits well known to those skilled in the art are not described in detail in order to highlight the main purpose of the present disclosure.

[0045] The experimental reagents, equipment, etc. involved in the present invention can be purchased from the market or prepared by yourself, and the present invention does not limit this. For some experimental steps, such as slicing operations, digital paddle scanning imaging and image processing, operations can be performed in combination with corresponding imaging equipment and image processing system software.

[0046] In this embodiment, the micro sample refers to animal cell tissue with a size of 0.5 mm.

[0047] Large samples refer to 5 mm comparison sample tissue, which can be understood in conjunction with the background description.

[0048] Example 1

[0049] This solution is to set a comparison sample for the tiny sample, slice it together with the tiny sample, form a glass slide and digitally scan it for imaging, so that the scanning device can locate, focus and shoot the tiny sample based on the comparison sample.

[0050] On the one hand, the present application provides a method for preparing a micro-sample pathology digital slide, comprising the following steps:

[0051] S1. Extract and pre-process tiny samples from animal model tissues;

[0052] S2, preparing a sample positioning marker and slicing it together with the micro-sample to form a pathology digital slide of the micro-sample;

[0053] S3, digitally scanning the pathology digital slide, positioning and focusing the micro-sample based on the sample positioning marker, and acquiring a WSI slice image of the micro-sample;

[0054] S4. Perform image processing on the WSI slice image, and locate the tiny sample based on the sample positioning marker.

[0055] This embodiment takes the preparation of ICR mouse liver tissue slides as an example.

[0056] Digital scanning was performed using the “Leica Aperio AT2 slide scanner” and scanning was performed using “20X scanning”.

[0057] The specific preparation steps are as follows:

[0058] 1. ICR mice were killed by cervical dislocation, the abdominal cavity was quickly opened, the left lobe of the liver was removed, and the liver was placed in 10% buffered neutral formalin solution and fixed overnight.

[0059] 2. Take the liver and cut it into liver tissue blocks with diameters of 0.5 mm (small sample) and 5 mm (large sample).

[0060] 3. Gradient dehydration: The tissue was immersed in gradient ethanol in sequence, with the concentrations of 60% ethanol, 70% ethanol, 80% ethanol, 90% ethanol, 95% ethanol I, 95% ethanol II, anhydrous ethanol I, and anhydrous ethanol II for 60 minutes each.

[0061] 4. Transparentization: After gradient dehydration, immerse the tissue in transparent agent I and transparent agent II for 20 minutes each.

[0062] 5 Wax immersion: After the tissue has become transparent, immerse it in paraffin wax I, paraffin wax II and paraffin wax III in turn for 40 minutes each.

[0063] 6. Paraffin embedding: Embed the micro-sample and the large sample side by side in the same wax block and slice them into 3μm sections.

[0064] 7.HE staining: bake at 65℃ for 10min, make xylene I and xylene II transparent for 8min respectively, treat with anhydrous ethanol I and anhydrous ethanol II for 5min respectively, treat with gradient alcohol 90% ethanol, 80% ethanol I, and 70% ethanol for 3min respectively, rinse with tap water for 3min, stain with hematoxylin staining solution for 7min, rinse with tap water for 3min, differentiate with 0.5﹪ hydrochloric acid ethanol for 8s, rinse with tap water for 3min, blue with 0.5﹪ dilute ammonia water for 1min, rinse with tap water for 2min, treat with 80% ethanol II and 95% ethanol for 3min respectively, stain with eosin staining solution for 1min, treat with 95% ethanol II and 95% ethanol III for 80s respectively, treat with anhydrous ethanol III and anhydrous ethanol IV for 5min respectively, use xylene III for 10min, seal the slides, and let them dry for later use.

[0065] Reagent preparation method:

[0066] Preparation of hematoxylin staining solution: first use 200 ml of ultrapure water to heat and dissolve potassium aluminum sulfate, then use 10 ml of anhydrous ethanol to dissolve hematoxylin and pour into the dissolved potassium aluminum sulfate solution and boil for 1 minute. After cooling slightly, slowly add 1 g of red mercuric oxide and continue heating and stirring until the dye solution turns dark purple. Cool with cold water, filter with filter paper and add 5 ml of acetic acid.

[0067] 0.5﹪ hydrochloric acid ethanol: Take 300ml of anhydrous ethanol, add it to 100ml of water, mix well, and you can get 75% ethanol. Take 2ml of concentrated hydrochloric acid, slowly add 398ml of 75% ethanol, mix well, and you are done.

[0068] 0.5﹪ dilute ammonia water: add 5 drops of concentrated ammonia water to 450ml of water and mix well.

[0069] 8. After drying, digital slides were scanned at 20X using a Leica Aper io AT2 slide scanner.

[0070] The scanning steps are as follows:

[0071] Instrument self-check: The slide scanner is turned on, self-checked, and the instrument is initialized until the LED screen displays "ScannerReady";

[0072] Prepare sections: Place sections on slide tray with coverslip up and label side out;

[0073] Setting parameters: Set the scanning magnification to 20X in the software;

[0074] Start scanning: Click Start, select Pre-Snapshot, adjust the scanning area frame to cover all tissues, and drag the green quadrilateral light spot in the control area to a clear area without tissues. At the same time, add focus on the tissues. The number of focus on large samples should be ≥4, and the number of focus on small samples should be ≥1;

[0075] Image processing: upload the captured images to the computer and use scanning software for image processing;

[0076] Save Image: Save the processed image for later analysis or printing.

[0077] 9. Results: Both large and small liver samples were successfully scanned.

[0078] like Figure 5 As shown, the low-magnification image of WS I shows a large sample (lower part of the image) and a small sample liver (upper part of the image), with clear boundaries and bright colors.

[0079] Depend on Figure 6 and Figure 7 The WSI high-power magnification image shown in the figure shows that the central vein of the liver of the large sample and the small sample is filled with red blood cells, the hepatocytes are well-formed, the nuclei are round, the nucleoli are obvious, and they are obviously eosinophilic, located in the center of the hepatocytes, and the cytoplasm is eosinophilic. The digital slide preparation effect of the liver pathology of the small sample is excellent.

[0080] Therefore, the present application solves the problem that it is difficult to locate and focus tiny samples under a slide scanner, and improves the slide preparation method to make the sample easier to identify and align on the slide. According to the characteristics of the tiny sample, select a suitable contrast enhancing substance. For cell-like tiny samples (such as organoid cells, cells in puncture objects), for example, when processing clinical puncture objects (tiny samples), if the puncture site is the liver, a small portion of normal animal liver tissue (large sample) can be selected. During the preparation process, the large sample and the tiny sample are placed side by side, and the large sample and the target tiny sample are used together to make the tiny sample clearly marked on the slide. The distribution of the sample on the slide is more conducive to scanning, reducing the scanning blind area caused by the sample being too small. At the same time, the problem of limited image information and background interference of the tiny sample is solved, and the clarity and recognition of the image are improved by adding appropriate contrast enhancing substances or optimizing the sample processing steps during the preparation process, so as to ensure that the pathological characteristics can be accurately presented. The large sample can be easily identified under the scanner by conventional staining or marking methods, so that the position of the tiny sample can be quickly determined by the relative relationship with the large sample, thereby achieving rapid positioning and accurate focusing.

[0081] The above-mentioned comprehensively improved slide preparation method can effectively solve the positioning and focusing problems of tiny samples under the slide scanner, reduce the scanning blind area, and improve the image quality, providing strong support for the pathological analysis of tiny samples.

[0082] It should be noted that, although animal liver is used as an example, those skilled in the art will understand that the present disclosure should not be limited thereto. In fact, the user can flexibly set the experimental object according to the actual application scenario, as long as the technical functions of the present application can be realized according to the above technology.

[0083] This embodiment uses a large sample to assist in locating a micro-sample. For example, normal animal liver tissue is used as a large sample in the processing of micro-samples of liver punctures. In addition to using a large sample of the same tissue, other samples with the same structure or properties as the micro-sample can also be selected as comparison markers. Choose different auxiliary sample types: but other known samples with similar characteristics or structures to the micro-sample can also be selected as auxiliary. For example, if the micro-sample is a trace sample of a special diseased cell, a cell sample with similar lesions but in a more typical and easier to identify stage can be selected as a reference for auxiliary positioning, which may be more conducive to comparing and locating the micro-sample. For example:

[0084] Preparation of digital slides of optic nerve of ICR mice

[0085] ICR mice were killed by cervical dislocation, the skull was quickly opened to remove the optic nerve, the abdominal cavity was opened to remove the left lobe of the liver, and placed in a 10% buffered neutral formalin solution and fixed overnight. The samples were collected the next day, the optic nerve was embedded as a whole, and another piece of liver was taken and placed in an embedding box together. Then it was dehydrated with 70%, 80%, 90%, 95%, and 100% ethanol solutions in sequence, and then placed in a xylene solution for transparency, then immersed in paraffin, and then embedded into paraffin blocks. 3μm sections were made and stained with hematoxylin-eosin. The sections were stained according to the present invention. The sections were digitally scanned and analyzed: The optic nerve of the ICR mouse is a bundle of nerve fibers connecting the eyes and the brain, mainly composed of the axons of retinal ganglion cells. After HE staining, the structure can be clearly displayed. Figure 8 As shown in the figure, the optic nerve is very small (about 230μm in diameter), and due to its own cell composition, the HE staining is very light, which is slightly difficult to distinguish with the naked eye. In contrast, the liver is brightly stained and has a clear contrast. Fig. 9 As shown, it is mainly composed of optic nerve fibers, optic nerve myelin, glial cells and blood vessels.

[0086] Therefore, the present invention uses a large sample to assist in positioning a tiny sample and clarifies the relative position of the large sample and the tiny sample (such as placement at a specific distance) to achieve auxiliary positioning. This method can effectively solve the problem that tiny samples are difficult to position and focus under a slide scanner, so that the tiny samples have clear marks on the slide, thereby achieving rapid positioning and accurate focusing. This method is innovative and practical, and may be a key part in the scope of protection of the invention, because it provides a simple and effective strategy to solve the problem of tiny sample positioning, which is different from the traditional method that only relies on the characteristics of the sample itself or complex instrument positioning.

[0087] Example 2

[0088] Based on the implementation principle of Example 1, the present application, on the other hand, proposes a method for preparing a micro-sample pathology digital slide, comprising the following steps:

[0089] S1. Extract and pre-process tiny samples from animal model tissues;

[0090] S2, slicing the tiny sample to form a pathology digital slide of the tiny sample, and adding a sample positioning marker on the pathology digital slide to provide a position marker for positioning and focusing the tiny sample on the pathology digital slide;

[0091] S3, digitally scanning the pathology digital slide, positioning and focusing the micro-sample based on the sample positioning marker, and acquiring a WSI slice image of the micro-sample;

[0092] S4. Perform image processing on the WSI slice image, and locate the tiny sample based on the sample positioning marker.

[0093] On the basis of Example 1, the present invention may also adopt other positioning marking methods. For example:

[0094] Use physical markers to assist positioning: In addition to relying on additional samples to assist positioning, you can also add some physical markers on the slide to help locate tiny samples. For example, set tiny metal markers or fluorescent markers at specific locations on the slide, and use the instrument to identify these markers to determine the approximate location range of the tiny sample, and then perform precise scanning and focusing. This method can avoid the interference that may be caused by introducing additional samples, and the position of the marker can be precisely designed and placed as needed.

[0095] Select other contrast enhancing substances: The invention mentions adding contrast enhancing substances to improve the clarity and recognition of the image. In addition to some existing chemical dyes and other substances, the use of new nanomaterials as contrast enhancers can also be explored. For example, some nanoparticles have unique optical properties and can produce strong scattering or fluorescence signals under light of a specific wavelength. When combined with a tiny sample, the contrast between the sample and the background can be significantly enhanced. Or use some specific antibodies combined with fluorescent markers to enhance the display of specific components, and select corresponding antibodies for labeling according to different pathological characteristics to improve the recognition of specific pathological structures in tiny samples.

[0096] The specific physical marker or type of contrast agent or contrast enhancement agent can be selected by the experimenter.

[0097] Therefore, the present invention adds appropriate contrast enhancement substances (such as selecting appropriate stains for different types of micro-samples) and optimizes the sample processing steps (such as fine control of washing, dehydration and other processes) during the preparation process to improve the clarity and recognition of the image and ensure that the pathological features can be accurately presented. In this way, the diagnostic difficulties caused by limited image information of micro-samples and background interference can be overcome, and there are significant technical improvements in the field of pathology slide preparation, providing technical support for the preparation of high-quality pathology digital slides.

[0098] Example 3

[0099] Embodiment 1 and Embodiment 2 can be combined to prepare a glass slide by integrating the two preparation modes to obtain a digital image with higher image clarity and micro-sample contrast.

[0100] Therefore, the present invention has the following technical advantages:

[0101] 1. Improve the accuracy of positioning and focusing:

[0102] Clearly identify the sample position: Due to its extremely small size, tiny samples are difficult to accurately locate during conventional slide preparation and scanning. The invented method can be used with the aid of large sample auxiliary positioning, slide markings or special slide materials to clearly mark tiny samples on the slide, so that the scanner can quickly and accurately locate and focus, reducing the risk of misdiagnosis or missed diagnosis due to positioning difficulties.

[0103] Reduce operational errors: It enables pathologists or researchers to more easily obtain clear images of tiny samples when using digital slides for analysis, improve the efficiency and accuracy of diagnosis and research, and reduce operational errors caused by difficulty in locating and focusing samples.

[0104] 2. Reduce scanning blind areas:

[0105] Evenly distribute samples: Tiny samples tend to be unevenly distributed on the slide, causing some areas to be missed during scanning, forming scanning blind spots. The improved preparation method can make the samples more evenly distributed on the slide, ensuring that the scanner can fully obtain the information of the samples, avoiding the impact of missing information on diagnosis and research results.

[0106] Multi-angle scanning coverage: For small samples with large numbers or complex distribution, multi-angle scanning can cover the samples more comprehensively, reduce information omissions caused by small and unevenly distributed samples, and improve the information integrity of digital slides.

[0107] 3. Improve image quality:

[0108] Enhance contrast effect: The image information of tiny samples is limited, and the contrast with the background may not be high. Adding contrast-enhancing substances or optimizing the sample processing steps can highlight the pathological characteristics of the sample, make the distinction between the sample and the background more obvious, and improve the clarity and recognition of the image.

[0109] 4. Facilitate data management and sharing:

[0110] Standardized preparation process: The invented method can standardize the preparation process of micro-sample pathology digital slides, so that the digital slides prepared by different laboratories or medical institutions are consistent, which is convenient for data management and comparison. This is of great significance for conducting multi-center research, establishing a pathology database, and realizing remote consultation.

[0111] Promote information sharing: High-quality digital slide images are easier to transmit and share, facilitating remote consultation and academic exchanges between pathologists, helping to promote the development of pathology and improve the efficiency of medical resource utilization.

[0112] The embodiments of the present disclosure have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or technical improvements in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A method for preparing a micro-sample pathology digital slide, characterized in that: The steps include: S1. Extract and pre-process tiny samples from animal model tissues; S2, preparing a sample positioning marker and slicing it together with the micro-sample to form a pathology digital slide of the micro-sample; S3, digitally scanning the pathology digital slide, positioning and focusing the micro-sample based on the sample positioning marker, and acquiring a WSI slice image of the micro-sample; S4. Perform image processing on the WSI slice image, and locate the tiny sample based on the sample positioning marker.

2. The method for preparing a micro-sample pathology digital slide according to claim 1, characterized in that: The sample positioning marker is a 5 mm large sample of the same tissue as the micro sample.

3. The method for preparing a micro-sample pathology digital slide according to claim 2, characterized in that: The micro-sample and the macro-sample are embedded in paraffin.

4. The method for preparing a micro-sample pathology digital slide according to claim 1, characterized in that: The sample positioning marker is another cell sample that is of a different tissue from the micro-sample but has the same or similar sample cell morphology characteristics.

5. The method for preparing a micro-sample pathology digital slide according to claim 1, characterized in that: The sample positioning marker is a contrast enhancement agent, which is used for component imaging of the tiny sample.

6. The method for preparing a micro-sample pathology digital slide according to claim 5, characterized in that: The contrast enhancing agent is one or more of the following contrast agents: Chemical stains, fluorescent contrast agents, or nanoenhancements.

7. A method for preparing a micro-sample pathology digital slide, characterized in that: The steps include: S1. Extract and pre-process tiny samples from animal model tissues; S2, slicing the tiny sample to form a pathology digital slide of the tiny sample, and adding a sample positioning marker on the pathology digital slide to provide a position marker for positioning and focusing the tiny sample on the pathology digital slide; S3, digitally scanning the pathology digital slide, positioning and focusing the micro-sample based on the sample positioning marker, and acquiring a WSI slice image of the micro-sample; S4. Perform image processing on the WSI slice image, and locate the tiny sample based on the sample positioning marker.

8. The method for preparing a micro-sample pathology digital slide according to claim 7, characterized in that: The sample positioning marker is a physical marker.

9. The method for preparing a micro-sample pathology digital slide according to claim 8, characterized in that: The physical marker is one or more of the following markers: Metallic or fluorescent markers.

Images