HE chromatin control, preparation method and application thereof

Abstract

The application provides an HE chromatin control product, a preparation method and application thereof, including a control slide, the control slide includes a first smear area and a second smear area, the first smear area and the second smear area are respectively arranged in different areas of the control slide, the first smear area is uniformly coated with mammalian peripheral blood mature red blood cells, and the second smear area is uniformly coated with mammalian peripheral blood lymphocytes. The HE chromatin control product provided by the application includes the combination of the mature red blood cell first smear area and the lymphocyte second smear area, can respectively and specifically reflect the staining effect of cytoplasm and nucleus in HE staining, realizes independent quality control of nucleus and plasma, the blood cell source is uniform, the difference of human body structure can be eliminated, batch uniformity and stability can be ensured, quantitative detection can be realized through a microspectrophotometer, objective and quantitative staining quality can be realized, the subjective and standardized difficulty problem of traditional quality control can be solved, and the accuracy and consistency of pathological quality control are improved.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to an HE staining quality control product, a method for preparing the HE staining quality control product, and its application. Background Technology

[0002] Hematoxylin-eosin (HE) staining is an indispensable routine staining technique in pathological diagnosis. It stains the cell nucleus blue and the cytoplasm red with hematoxylin and eosin respectively, providing a basis for pathological morphological observation and disease diagnosis. Currently, HE staining quality control commonly uses tissue microarray sections made from placental, appendix, and colon tissues. After staining, the staining effect is manually interpreted by professionals. While this method is a routine quality control approach in the industry, it has significant limitations: the interpretation results are highly dependent on the operator's professional experience, easily influenced by subjective visual judgment, and lack quantification, objectivity, and stability. It is difficult to achieve standardized and quantitative quality evaluation, failing to meet the long-term stable and controllable requirements of clinical pathological testing for HE staining. Furthermore, it hinders the unified comparison of HE staining quality control results between laboratories and the subsequent promotion and application of artificial intelligence in pathological diagnosis. Summary of the Invention

[0003] To address the problems of existing HE staining quality control methods relying on subjective human judgment, resulting in poor objectivity and difficulty in achieving quantitative, stable, and standardized quality control, this paper provides an HE staining quality control product, its preparation method, and its application.

[0004] The technical solution adopted by the present invention to solve the above-mentioned technical problems is as follows: On one hand, the present invention provides an HE staining quality control material, including a first smear area and a second smear area, wherein the first smear area and the second smear area are disposed on the same glass slide, wherein the first smear area is uniformly coated with mature erythrocytes from mammalian peripheral blood, and the second smear area is uniformly coated with lymphocytes from mammalian peripheral blood.

[0005] Optionally, the mature erythrocytes of mammalian peripheral blood in the first smear area are uniformly distributed in a monolayer, and the number of mature erythrocytes of mammalian peripheral blood per unit area is 1.4 × 10⁻⁶. 6 pcs / cm 2 ~1.5×10 6 pcs / cm 2 .

[0006] Optionally, the mammalian peripheral blood lymphocytes in the second smear area are uniformly distributed in a monolayer, and the number of mammalian peripheral blood lymphocytes per unit area is 1.4 × 10⁻⁶. 6 pcs / cm 2 ~1.5×10 6 pcs / cm2 .

[0007] Optionally, the method for preparing the HE staining quality control includes the following steps: Collect mammalian blood, add an anticoagulant, and set aside for later use; The separation solution was added to mammalian blood, and further centrifugation was performed to obtain mature erythrocytes and lymphocytes from mammalian peripheral blood. The cells were then resuspended in physiological saline to obtain a cell suspension. Mature erythrocytes from mammalian peripheral blood were added to the first smear area and allowed to settle. Lymphocytes from mammalian peripheral blood were added to the second smear area and allowed to settle. After settling, the cells were fixed with glutaraldehyde solution to obtain the HE staining quality control.

[0008] On the other hand, the HE staining quality control product provided in this application, or the HE staining quality control product prepared by the method for preparing the HE staining quality control product, is used in HE staining quality control.

[0009] Optionally, the application of the HE staining quality control material in HE staining quality control includes the following operations: HE staining quality control samples and ex vivo tissue samples to be tested were placed in the same environment and HE staining was performed simultaneously. After staining, the absorbance values ​​of the first smear region and the second smear region in the HE staining quality control material were detected by a micro spectrophotometer. The absorbance values ​​of the first and second smears are compared with the preset reference absorbance value range. If the absorbance value is within the reference absorbance value range, the HE staining quality is deemed acceptable; otherwise, it is deemed unacceptable.

[0010] Optionally, in the application of the HE staining quality control material in HE staining quality control, the acceptable range of the reference absorbance value is determined by the following method: After HE staining of the standard HE quality control tissue microarray, multiple professional pathologists judged the HE staining effect. To ensure the best staining effect, 20 HE staining quality control slides were stained with HE. The absorbance values ​​of the first smear region and the second smear region were measured by a microspectrophotometer. The mean and standard deviation of the absorbance values ​​of the first smear region and the second smear region were calculated, and the result of the mean ± 5.5 times the standard deviation was used as the reference value for the quality control of eosin staining and hematoxylin staining, respectively.

[0011] The beneficial effects of this application are as follows: The HE staining quality control provided in this application includes a first smear area and a second smear area, both positioned on the same glass slide. Mature erythrocytes from mammalian peripheral blood are uniformly smeared in the first smear area, and mammalian lymphocytes are uniformly smeared in the second smear area. This HE staining quality control uses a dual-smear setup, separately fixing mature erythrocytes and lymphocytes from mammalian peripheral blood. Anucleated erythrocytes are stained with eosin, specifically reflecting the concentration, uniformity, and stability of cytoplasmic staining. Nucleated lymphocytes are precisely stained with hematoxylin, specifically reflecting the intensity, specificity, and consistency of nuclear staining, achieving independent quality control of nuclear and cytoplasmic staining effects. The mammalian blood cells are of uniform origin, with highly consistent cell morphology and composition, eliminating differences in human tissue donors and cell density from the source. To address batch-to-batch baseline deviations caused by variations in homogeneity and nucleoplasmic ratio, the homogeneity and repeatability of the quality control materials are ensured. Furthermore, relying on the biological characteristics of mature erythrocytes exhibiting eosin staining (red or light red) and lymphocytes being stained with hematoxylin (blue), a stable and uniform objective reference carrier can be provided for HE staining reactions, further avoiding the subjectivity and experience dependence brought about by manual interpretation. The two types of standardized cell smears can be adapted to a microspectrophotometer for quantitative absorbance detection, converting the staining effect into quantifiable values, and realizing the objective measurement of key indicators such as staining intensity and uniformity. This effectively solves the technical problems of strong subjectivity, insufficient objectivity, and difficulty in stable standardization in existing HE staining quality control, significantly improving the accuracy and consistency of staining quality control, and thus meeting the application needs of clinical pathological quantitative quality control and AI diagnosis. Attached Figure Description

[0012] Figure 1 This is a staining diagram of the appendix, placenta, and colon tissue provided in Comparative Example 1. Detailed Implementation

[0013] To make the technical problems solved, the technical solutions, and the beneficial effects of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0014] This invention provides a HE staining quality control, comprising a control slide. The control slide includes a first smear region and a second smear region, which are respectively located in different areas of the control slide. Mature erythrocytes from mammalian peripheral blood are uniformly smeared in the first smear region, and mammalian peripheral blood lymphocytes are uniformly smeared in the second smear region. In specific implementation, it should be noted that hematoxylin and eosin are used as the main dyes in the HE staining quality control, which can stain the cell nucleus and cytoplasm, respectively. Hematoxylin is a basic dye that stains the cell nucleus and ribosomes blue, while eosin is an acidic dye that stains the cytoplasm red or pale red.

[0015] Specifically, the HE staining quality control material approved in this application simultaneously sets up a first smear area and a second smear area in different regions of the quality control slide. The first smear area is uniformly coated with mature mammalian peripheral blood erythrocytes, and the second smear area is uniformly coated with mammalian peripheral blood lymphocytes. This HE staining quality control material uses independent dual-smear areas to fix mature mammalian peripheral blood erythrocytes and lymphocytes respectively. Anucleated erythrocytes are stained with eosin, which specifically reflects the concentration, uniformity, and stability of cytoplasmic staining. Nucleated lymphocytes are precisely stained with hematoxylin, which specifically reflects the intensity, specificity, and consistency of nuclear staining, achieving independent quality control of nuclear and cytoplasmic staining effects. The mammalian blood cells are of uniform origin, with highly consistent cell morphology and composition, eliminating differences in human tissue donors and cell density from the source. To address batch-to-batch baseline deviations caused by variations in homogeneity and nucleoplasmic ratio, the quality control materials themselves are made uniform and reproducible. Furthermore, relying on the biological characteristics of mature erythrocytes exhibiting eosin staining (red or light red) and lymphocytes being stained with hematoxylin (blue), a stable and uniform objective reference carrier can be provided for HE staining reactions, further avoiding the subjectivity and experience dependence brought about by manual interpretation. The two types of standardized cell smears can be adapted to a microspectrophotometer for quantitative absorbance detection, converting the staining effect into quantifiable values, and realizing the objective measurement of key indicators such as staining intensity and uniformity. This effectively solves the technical problems of strong subjectivity, insufficient objectivity, and difficulty in stable standardization in existing HE staining quality control, significantly improving the accuracy and consistency of staining quality control, and thus meeting the application needs of quantitative quality control and diagnosis in clinical pathology.

[0016] Specifically, mature erythrocytes lack nuclei and organelles, containing only cytoplasm. During HE staining, they bind only to eosin, completely eliminating nuclear staining interference. This provides a specific and precise reflection of the staining intensity, uniformity, and stability of eosin on the cytoplasm, offering a single, pure quality control target for the cytoplasmic staining stage in HE staining. Mature erythrocytes are morphologically uniform, regularly sized, and easily standardized in origin. They can be thoroughly mixed and spread onto a batch of slides, avoiding baseline deviations caused by differences in cell density and composition in human tissues, ensuring the uniformity of quality control samples within a batch. Furthermore, their staining is singular and stable, allowing for quantitative absorbance detection using a microspectrophotometer. This provides an objective and quantifiable evaluation basis for eosin staining quality, improving the accuracy and repeatability of HE staining cytoplasmic quality control.

[0017] In some embodiments, the mammalian peripheral blood mature erythrocytes in the first smear region are uniformly distributed in a monolayer, and the number of mammalian peripheral blood mature erythrocytes per unit area is 1.4 × 10⁻⁶. 6 pcs / cm 2 ~1.5×10 6 pcs / cm 2 .

[0018] Mature erythrocytes from mammalian peripheral blood in the first smear region were arranged in a monolayer with uniform distribution, and the number of mature erythrocytes from mammalian peripheral blood per unit area was further limited to 1.4 × 10⁻⁶. 6 pcs / cm 2 ~1.5×10 6 pcs / cm 2 Within a specific range, it can significantly improve the stability and quantitative accuracy of HE staining quality control from the perspectives of staining and optical detection. Specifically, the uniform distribution of a single layer can avoid localized excessive staining, uneven light transmission, and absorbance detection deviations caused by cell overlap and accumulation, ensuring that mature red blood cells and eosin dye fully and uniformly combine, so that the cytoplasmic staining intensity truly reflects the state of the staining system. At the same time, the constant number of cells per unit area can ensure that the total amount of staining substrate in each batch of quality control smears is consistent, so that the absorbance value and staining intensity remain stable, reducing the differences between batches of quality control smears, providing a quantitative, repeatable, and standardized objective quality control basis for HE staining, and significantly improving the reliability and consistency of quality control results.

[0019] Specifically, lymphocytes in the peripheral blood of mammals can stably exhibit basophilic deep blue staining of nuclear hematoxylin in HE staining, providing a relatively standard reference for nuclear and plasma staining in HE staining; Using lymphocytes as quality control cells can avoid the interpretation interference caused by the complex cell types and disordered structures in tissue sections. At the same time, combined with microspectrophotometry, stable and reliable absorbance values ​​can be obtained, which improves the quantitative accuracy and standardization of HE staining quality control.

[0020] In some embodiments, the mammalian peripheral blood lymphocytes in the second smear region are uniformly distributed in a monolayer, and the number of mammalian peripheral blood lymphocytes per unit area is 1.4 × 10⁻⁶. 6 pcs / cm 2 ~1.5×10 6 pcs / cm 2 .

[0021] Setting the mammalian peripheral blood lymphocytes in the second smear region to a monolayer with uniform distribution and limiting the number of cells per unit area significantly improves the stability and quantitative accuracy of HE staining quality control in terms of staining and optical detection. Specifically, the monolayer uniform distribution avoids uneven staining, excessive local staining, or abnormal transmittance caused by cell overlap and accumulation, ensuring that each lymphocyte fully and uniformly binds to the hematoxylin dye, truly reflecting the actual staining effect. At the same time, the constant number of cells per unit area ensures that the total amount of staining substrate is consistent in each batch of quality control smears, keeping the absorbance value and staining intensity stable, reducing the difference between batches of quality control smears, thereby effectively reducing the error caused by subjective human interpretation, providing a quantitative, repeatable, and standardized objective quality control basis for HE staining, and significantly improving the reliability and consistency of quality control results.

[0022] In some embodiments, the quality control slides may be of one type: cationic slides, silanized slides, and poly-L-lysine slides.

[0023] Selecting slides with good cell adhesion properties can provide a stable and reliable carrier support for HE staining quality control from the perspective of material-cell binding mechanism. Specifically, such slides can physically adsorb and firmly fix mature erythrocytes and lymphocytes from mammalian peripheral blood onto the slide surface, avoiding cell detachment, displacement, or distribution changes during operation. This ensures that mature erythrocytes and lymphocytes always maintain a uniform monolayer distribution, making the staining reaction sufficient and uniform, thereby ensuring that the absorbance values ​​detected by the spectrophotometer are stable, accurate, and repeatable.

[0024] In some embodiments, the method for preparing the HE staining quality control includes the following steps: Collect mammalian blood, add an anticoagulant for later use, and separate mature red blood cells and lymphocytes within 2 hours; Add mammalian blood to the separation solution according to the instructions for use of the lymphocyte separation solution. Centrifuge to obtain lymphocytes. Wash with physiological saline and resuspend in physiological saline to adjust the cell concentration to obtain a lymphocyte suspension. Take an appropriate amount of anticoagulated whole blood, centrifuge it, collect mature red blood cells, wash them with physiological saline, resuspend them in physiological saline and adjust the cell concentration to obtain a red blood cell suspension. Mature erythrocyte suspension from mammalian peripheral blood was settled in the first smear area for 30 minutes to form the first smear area. Similarly, a suspension of lymphocytes from mammalian peripheral blood was settled in the second area for 30 minutes to form the second smear area. The smears were then fixed with 2.5% glutaraldehyde solution pre-cooled to 4°C for 2 hours to obtain the HE staining quality control.

[0025] The preparation method of the HE staining quality control material specifically includes the following steps: Collect mammalian blood, add citrate anticoagulant for later use; Add mammalian blood to the separation solution according to the instructions for use of the lymphocyte separation solution. Centrifuge to obtain lymphocytes according to the instructions for use of the lymphocyte separation solution. After washing with physiological saline, resuspend in physiological saline and adjust the cell concentration to obtain a lymphocyte suspension. Take an appropriate amount of anticoagulated whole blood, centrifuge it, collect mature red blood cells, wash them with physiological saline, resuspend them with physiological saline and adjust the cell concentration to obtain a red blood cell suspension. Mature erythrocyte suspension from mammalian peripheral blood was settled in the first smear area for 30 minutes to form the first smear area. Similarly, a suspension of lymphocytes from mammalian peripheral blood was settled in the second area for 30 minutes to form the second smear area. The samples were then fixed with 2.5% glutaraldehyde solution pre-cooled to 4°C for 2 hours to obtain the quality control sample.

[0026] Specifically, the centrifugation process includes: After adding anticoagulated whole blood to the lymphocyte separation medium, centrifuge at 800g for 30 minutes, carefully aspirate the lymphocyte layer, wash with physiological saline, resuspend in physiological saline, and adjust the cell concentration.

[0027] Take an appropriate amount of anticoagulated whole blood, centrifuge at 500g for 5 minutes, carefully aspirate the mature red blood cells from the bottom, wash with physiological saline, resuspend in physiological saline, and adjust the cell concentration.

[0028] In some embodiments, the method for preparing the HE staining quality control product includes a citrate anticoagulant.

[0029] The separation solution is a lymphocyte separation solution, and the fixative is a 2.5% glutaraldehyde solution pre-cooled at 4°C.

[0030] In another embodiment of this application, the HE staining quality control product, or the HE staining quality control product prepared by the method described above, is used in HE staining quality control.

[0031] Specifically, this quality control material uses mature erythrocytes isolated from mammalian peripheral blood that specifically respond to eosin staining and lymphocytes that specifically respond to hematoxylin staining. It can characterize the staining intensity and specificity of hematoxylin and eosin, respectively. By using cell suspensions to produce HE staining quality control materials in batches, the uniformity and stability of the quality control materials are ensured. The smear cells are uniformly distributed and the color signal is pure. It can be adapted to a microspectrophotometer to realize quantitative absorbance detection, transforming subjective human judgment into objective values. It realizes quantitative, standardized, and traceable HE staining quality evaluation, which can not only monitor the stability of the staining process in real time, but also be used for inter-laboratory quality comparison and the consistency requirements of staining in pathological diagnosis, thereby improving the objectivity, accuracy and universality of HE staining quality control.

[0032] In another embodiment of this application, the application of the HE staining quality control material in HE staining quality control includes the following operations: HE staining quality control samples and ex vivo tissue samples to be tested were placed in the same environment and HE staining was performed simultaneously. After staining, the absorbance values ​​of the first and second smear regions in the HE staining quality control were measured using a micro spectrophotometer. The absorbance values ​​of the first and second smear areas were compared with reference values. If the values ​​were within the reference range, the HE staining quality was considered to be under control; otherwise, the staining quality was considered to be unqualified.

[0033] Specifically, under the same batch, conditions, and conditions as the tissue to be tested, the quality control samples are stained synchronously to ensure that they have completely consistent temperature, time, staining concentration, and differentiation process with the clinical samples, so as to truly reflect the actual staining state of the samples to be tested. The absorbance of the first smear area (red blood cells) and the second smear area (lymphocytes) are detected by a microspectrophotometer, which can convert the results of eosin staining and hematoxylin staining into quantitative values, eliminating the subjectivity and experience dependence of manual interpretation. The measured absorbance is compared with the preset qualified range to realize the quantification and repeatability of HE staining quality, and ensure the stability and consistency of basic staining for pathological diagnosis.

[0034] In another embodiment of this application, the HE staining quality control material is used in HE staining quality control, and the reference absorbance value is determined by the following method: After HE staining of the standard HE quality control tissue microarray, the HE staining effect was judged by multiple professional pathologists. When the staining effect was optimal, 20 HE staining quality control slides were stained with HE. The absorbance values ​​of the first smear region and the second smear region were measured by a microspectrophotometer. The mean and standard deviation (s) of the absorbance values ​​of the first smear region and the second smear region were calculated, and the mean ± 5.5s of the two values ​​were used as the reference values ​​for eosin staining and hematoxylin staining quality control, respectively.

[0035] It should be noted that in the quality control testing of medical experiments, when establishing the initial quality control range and setting the target value and standard deviation, it is usually necessary to conduct no less than 20 repeated tests. This is because single or small number of test results are easily affected by random errors and the data is not representative enough. However, 20 or more independent repeated tests can fully reflect the random fluctuations of the testing system, reduce the deviation caused by accidental errors, and make the calculated parameters such as mean, standard deviation, and coefficient of variation more consistent with the true performance of the testing system. This can ensure the accuracy and effectiveness of subsequent quality control results. Based on this condition, HE staining operation was also performed on 20 HE staining quality control samples in this application.

[0036] The present invention will be further illustrated by the following examples.

[0037] Example 1 This embodiment illustrates the HE staining quality control material and its preparation method disclosed in this invention, including the following steps: Collect mammalian blood, add an anticoagulant for later use; Anticoagulated whole blood was carefully added to the upper layer of the separation solution. The lymphocytes were centrifuged according to the instructions of the lymphocyte separation solution to obtain mammalian peripheral blood lymphocytes. After washing with physiological saline, physiological saline was added to resuspend the cells and the cell concentration was adjusted to obtain the lymphocyte suspension. In addition, a portion of anticoagulated mammalian whole blood was centrifuged to obtain mature mammalian red blood cells. After washing with physiological saline, the cells were resuspended in physiological saline and the cell concentration was adjusted to obtain a red blood cell suspension. Add 300 μL of mature mammalian peripheral blood erythrocyte suspension to the first smear area and allow it to settle fully to form the first smear area. Add 300 μL of mammalian peripheral blood lymphocyte suspension to the second smear area and allow it to settle fully to form the second smear area. Then fix the cells in the smear areas with 2.5% glutaraldehyde solution pre-cooled to 4°C for 2 hours to obtain the quality control sample.

[0038] dyeing: Wash the smear with water for 30 seconds, then immerse it in hematoxylin for 6 minutes, wash it with water for 2 minutes, differentiate it for 12 seconds, wash it with water for 30 minutes, immerse it in the blueing solution for 1 minute, wash it with water for 2 minutes, wash it with 95% ethanol for 10 seconds, immerse it in eosinol solution for 40 seconds, differentiate it twice with 95% ethanol (20 seconds each time), dehydrate it with graded ethanol, clear it with xylene, and mount it. Detection: After staining, the absorbance of the smears was measured using a microspectrophotometer. The results are as follows: The maximum absorption peak in the first smear region (mature red blood cells) is located at 533 nm, with an absorbance value of 0.72. The maximum absorption peak in the second smear region (lymphocytes) is located at 600 nm, with an absorbance value of 0.28. In Example 1, after standardized HE staining using the HE staining quality control material prepared according to the present invention, the absorbance of mature erythrocytes in the first smear region was 0.72 at the maximum absorption peak of 533 nm, which can specifically and stably reflect the staining effect of eosin cytoplasm. The absorbance of lymphocytes in the second smear region was 0.28 at the maximum absorption peak of 600 nm, which can accurately characterize the hematoxylin nuclear staining state. The values ​​are stable and highly specific, and independent quantitative quality control of nuclear and cytoplasmic staining can be achieved.

[0039] Comparative Example 1 This comparative example used regular tissue microarray HE staining, including the following procedures: Appendix, placenta, and colon tissues were embedded into tissue microarrays. After sectioning and baking, the sections were dewaxed with xylene, then dexylene was removed to water using a gradient of ethanol and water. Finally, the sections were stained with hematoxylin and eosin (HE) according to the method for smear staining. The staining results are shown in the figure. Figure 1 .

[0040] Comparative Example 1 uses traditional staining techniques to perform HE staining on tissue microarrays of appendix, placenta, and colon. This method is easily affected by factors such as differences in human tissue donors, mixed cell types, uneven nucleo-cytoplasmic ratio, and fluctuations in cell density. The staining results are prone to batch-to-batch variations and can only be interpreted subjectively by pathologists. It cannot be converted into quantitative absorbance values, and has the defects of strong subjectivity, poor standardization, and poor repeatability. The comparison highlights that the HE staining quality control provided in this application can effectively achieve objective quantification of staining effects and batch-to-batch uniformity and stability, thereby improving the accuracy and standardization of HE staining pathological quality control and avoiding the drawbacks of traditional tissue microarray quality control.

[0041] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A HE staining quality control product, characterized in that, The system includes a quality control slide, which comprises a first smear region and a second smear region. The first smear region and the second smear region are respectively located in different areas of the quality control slide. Mature erythrocytes from mammalian peripheral blood are uniformly coated in the first smear region, and mammalian peripheral blood lymphocytes are uniformly coated in the second smear region.

2. The HE staining quality control product according to claim 1, characterized in that, In the first smear area, mature mammalian peripheral blood erythrocytes are uniformly distributed in a monolayer, and the number of mature mammalian peripheral blood erythrocytes per unit area is 1.4 × 10⁻⁶. 6 pcs / cm 2 ~1.5×10 6 pcs / cm 2 .

3. The HE staining quality control product according to claim 1, characterized in that, In the second smear area, mammalian peripheral blood lymphocytes were uniformly distributed in a monolayer, and the number of mammalian peripheral blood lymphocytes per unit area was 1.4 × 10⁻⁶. 6 pcs / cm 2 ~1.5×10 6 pcs / cm 2 .

4. The method for preparing HE staining quality control material according to any one of claims 1 to 3, characterized in that, Includes the following steps: Collect mammalian blood and add an anticoagulant for later use; The separation solution was added to mammalian blood, and further centrifugation was performed to obtain mature erythrocytes and lymphocytes from mammalian peripheral blood. The cells were then resuspended in physiological saline to obtain a cell suspension. Mature erythrocytes from mammalian peripheral blood were added to the first smear area and allowed to settle. Lymphocytes from mammalian peripheral blood were added to the second smear area and allowed to settle. After settling, the cells were fixed with glutaraldehyde solution to obtain the HE staining quality control.

5. The application of an HE staining quality control product as described in any one of claims 1 to 3, or an HE staining quality control product prepared by the method for preparing the HE staining quality control product as described in claim 4, in HE staining quality control.

6. The application of the HE staining quality control material according to claim 5 in HE staining quality control, characterized in that, Includes the following operations: HE staining quality control samples and ex vivo tissue samples to be tested were placed in the same environment and HE staining was performed simultaneously. After staining, the absorbance values ​​of the first smear region and the second smear region in the HE staining quality control material were detected by a micro spectrophotometer. The absorbance values ​​of the first and second smears are compared with the preset reference absorbance value range. If the absorbance value is within the reference absorbance value range, the HE staining quality is deemed acceptable; otherwise, it is deemed unacceptable.

7. The application of the HE staining quality control material according to claim 6 in HE staining quality control, characterized in that, The acceptable range of the reference absorbance value is determined by the following method: After HE staining of the standard HE quality control tissue microarray, multiple professional pathologists judged the HE staining effect. To ensure the best staining effect, 20 HE staining quality control slides were stained with HE. The absorbance values ​​of the first smear region and the second smear region were measured by a microspectrophotometer. The mean and standard deviation of the absorbance values ​​of the first smear region and the second smear region were calculated, and the result of the mean ± 5.5 times the standard deviation was used as the reference value for the quality control of eosin staining and hematoxylin staining, respectively.

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