An immunohistochemical staining method and apparatus

Abstract

An immunohistochemical staining method and apparatus, the method comprising: a cleaning height calculation step of placing a slide carrying a sample after antigen repair in a water tank such that the slide is in a water bath, calculating the optimal cleaning height of the slide, and setting parameters according to the optimal cleaning height; a cleaning step of controlling a pipetting arm to aspirate a reagent, dispense it onto each slide, and after the reaction is completed, aspirate a cleaning agent and clean the slide according to the set parameters to obtain a slide after immunohistochemical staining. The present invention effectively avoids dry slides, has a simple experimental process, and uses a small amount of reagents.

Description

Technical Field

[0001] The present invention relates to the technical field of immunohistochemical staining, and particularly relates to an immunohistochemical staining method and device. Background Art

[0002] During the use of an immunohistochemical staining machine, the technical conditions that must be ensured are that the glass slide does not dry during the staining process to produce non-specific background; hematoxylin is not overstained to interfere with the color development of positive samples; problems such as sample blank in the central area of the sample, etc. In addition, when using the staining machine to stain the same batch, it is necessary to ensure the uniformity of the staining effect on the glass slides, otherwise it may affect the accuracy of the staining result.

[0003] In the existing staining methods, after the antigen retrieval is completed, if the water bath temperature is too high, it is easy to cause the glass slide to dry. In the staining experiment, the non-uniformity of the temperature will make the staining effect of the glass slides non-uniform and unstable. When using an air pump to clean the glass slides, the samples in the central area of the glass slides will also be sucked away by the air pump, resulting in a phenomenon of central blank in the staining result. When sucking, dispensing and mixing the reagents, the reagent sucking accuracy is insufficient, and there is a phenomenon of pure water hanging on the pipette needle wall after cleaning, which has a negative impact on reagent contamination and dilution.

[0004] Some existing technologies adopt adding a small plastic cover on the glass surface to provide a closed incubation environment for the immune reaction and prevent the volatilization of reagents, such as the bond-max of Leica. This solution has high requirements for temperature control accuracy, otherwise there is still a high possibility of dry slides.

[0005] There are also other staining machines on the market developed by imitating the manual staining method. The experimental process is relatively cumbersome, requiring multiple transfers of the glass slides. The instrument structure is relatively complex and the reagent consumption is large. Summary of the Invention

[0006] According to a first aspect, in one embodiment, an immunohistochemical staining method is provided, including:

[0007] A cleaning height calculation step of placing a glass slide carrying a sample after antigen retrieval in a water bath, calculating the optimal cleaning height of the glass slide, and setting parameters according to the optimal cleaning height;

[0008] A cleaning step including controlling a pipetting arm to suck reagents and dispense them onto each glass slide. After the reaction is completed, sucking a cleaning agent and cleaning the glass slides according to the set parameters to obtain an immunohistochemically stained glass slide.

[0009] According to a second aspect, in one embodiment, an immunohistochemical staining device is provided, including:

[0010] A cleaning height calculation module places the slide carrying the sample after antigen repair in a water tank so that the slide is in a water bath, calculates the optimal cleaning height of the slide, and sets parameters according to the optimal cleaning height;

[0011] A cleaning module includes a controlled pipetting arm that aspirates reagents and distributes them onto each slide. After the reaction is completed, it aspirates a cleaning agent and cleans the slide according to the set parameters to obtain a slide after immunohistochemical staining.

[0012] According to a third aspect, in an embodiment, an immunohistochemical staining device is provided, including:

[0013] A memory for storing programs;

[0014] A processor for implementing the method according to any one of the first aspects by executing the programs stored in the memory.

[0015] According to a fourth aspect, in an embodiment, a computer-readable storage medium is provided, on which a program is stored, and the program can be executed by a processor to implement the method according to any one of the first aspects.

[0016] According to an immunohistochemical staining method and device of the above embodiments, the present invention effectively avoids dry slides, the experimental process is simple, and the reagent consumption is small. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 It is a flowchart for controlling a staining machine to perform cell staining. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0018] The present invention will be further described in detail below in conjunction with the drawings through specific embodiments. In the following embodiments, many details are described to enable a better understanding of the present application. However, those skilled in the art can easily recognize that some of the features can be omitted in different situations or replaced by other materials and methods. In some cases, some operations related to the present application are not shown or described in the specification to avoid overwhelming the core part of the present application with excessive description. For those skilled in the art, it is not necessary to describe these related operations in detail, and they can fully understand the related operations based on the description in the specification and general technical knowledge in the art.

[0019] In addition, the features, operations, or characteristics described in the specification can be combined in any suitable manner to form various embodiments. At the same time, the steps or actions in the method description can also be reordered or adjusted in a manner that is obvious to those skilled in the art. Therefore, the various sequences in the specification and drawings are only for clearly describing a certain embodiment and do not mean a necessary sequence, unless it is stated that a certain sequence must be followed.

[0020] The serial numbers assigned to the components herein, such as "first", "second", etc., are only used to distinguish the objects described and do not have any sequential or technical meaning.

[0021] According to a first aspect, in one embodiment, there is provided an immunohistochemical staining method, comprising:

[0022] A cleaning height calculation step of placing the slide carrying the sample after antigen repair in a water tank such that the slide is in a water bath, calculating the optimal cleaning height of the slide, and setting parameters according to the optimal cleaning height;

[0023] A cleaning step including controlling a pipetting arm to aspirate a reagent, dispense it onto each slide, and after the reaction is completed, aspirate a cleaning agent and clean the slide according to the set parameters to obtain a slide after immunohistochemical staining.

[0024] In one embodiment, the cleaning step includes a first round of cleaning, a second round of cleaning, a third round of cleaning, a fourth round of cleaning, a fifth round of cleaning, a sixth round of cleaning, a seventh round of cleaning, and an eighth round of cleaning.

[0025] In one embodiment, the seventh round of cleaning includes: controlling the pipetting arm to aspirate a staining agent and dispense it onto each slide, waiting for a certain time after the dispensing of the slides in one reaction water tank is completed, and then cleaning the slides with a cleaning agent according to the set parameters, and dispensing the staining agent and cleaning the slides in other water tanks in the same manner.

[0026] In one embodiment, the certain time is 0.5 - 2 s, preferably 1 s.

[0027] In one embodiment, the staining agent includes hematoxylin.

[0028] In one embodiment, the cleaning agent used in the seventh round of cleaning contains water.

[0029] In one embodiment, in the cleaning height calculation step, the lower surface of the slide is in the water bath while the sample area on the upper surface is not immersed in the water bath, and then the optimal cleaning height of the slide is calculated.

[0030] In one embodiment, according to the set parameters, control the pipetting arm to clean the slide to obtain a slide after immunohistochemical staining.

[0031] In one embodiment, in the cleaning step, for the second round and each subsequent round of cleaning, before each round of cleaning, the pipetting arm is controlled to aspirate the required reagent to be added and dispense it onto the glass slide. After waiting for the reaction of the reagent to be completed, the glass slide is cleaned with a cleaning agent according to the set parameters.

[0032] In one embodiment, in the cleaning step, before the second round of cleaning, the reagent added is the DS-A reagent.

[0033] In one embodiment, in the cleaning step, before the third round of cleaning, the reagent added is the DS-B reagent.

[0034] In one embodiment, in the cleaning step, before the fourth round of cleaning, the reagent added is the DS-C reagent.

[0035] In one embodiment, in the cleaning step, before the fifth round of cleaning, the reagent added is the DAB reagent.

[0036] In one embodiment, the DAB reagent contains the DS-D and DS-E reagents.

[0037] In one embodiment, in the cleaning step, before the sixth round of cleaning, the reagent added is the Fast-Red reagent.

[0038] In one embodiment, the Fast-Red reagent contains the DS-F and DS-G reagents.

[0039] In one embodiment, in the DAB reagent, by volume, DS-D:DS-E = 1:33, and this ratio is denoted as d;

[0040] In the Fast-Red reagent, by volume, DS-F:DS-G = 1:50, and this ratio is denoted as f;

[0041] The total amount of reagents required for DAB and Fast-Red is denoted as S;

[0042] Volume of DS-D = [S × d] + 1;

[0043] Volume of DS-E = Volume of DS-D × 33;

[0044] Volume of DS-F = [S × f] + 1;

[0045] Volume of DS-G = Volume of DS-D × 55.

[0046] The volume is the volume of the corresponding reagent.

[0047] By the above calculation method, the problem of inaccurate reagent mixing ratio caused by the pipetting needle's inability to achieve a liquid absorption resolution accuracy of 0.1 level due to the calculated value being a decimal type data is avoided.

[0048] In one embodiment, the cleaning agent used in the first to sixth rounds of cleaning contains PBST.

[0049] In one embodiment, in the cleaning step, before the eighth round of cleaning, the added reagent is the blue-returning solution.

[0050] In one embodiment, in the cleaning step, the cleaning agent used in the eighth round of cleaning contains water.

[0051] In one embodiment, in the cleaning step, the experimental platform for placing the glass slides includes at least one area, and the glass slides in each area are stained separately.

[0052] In one embodiment, in the cleaning step, the number of the areas is 1 - 10, preferably 6.

[0053] In one embodiment, in the cleaning step, when the pipetting arm is flushed and cleaned in pure water, the pipetting needle is slowly lifted.

[0054] According to a second aspect, in one embodiment, there is provided an immunohistochemical staining device, including:

[0055] A cleaning height calculation module, which places the glass slide carrying the sample after antigen repair in a water tank so that the glass slide is in a water bath, calculates the optimal cleaning height of the glass slide, and sets parameters according to the optimal cleaning height;

[0056] A cleaning module, including controlling a pipetting arm, sucking reagents, distributing them onto each glass slide, and after the reaction is completed, sucking the cleaning agent and cleaning the glass slides according to the set parameters to obtain glass slides after immunohistochemical staining.

[0057] According to a third aspect, in one embodiment, there is provided an immunohistochemical staining device, including:

[0058] A memory for storing programs;

[0059] A processor for implementing the method according to any one of the first aspects by executing the programs stored in the memory.

[0060] According to a fourth aspect, in one embodiment, there is provided a computer-readable storage medium, on which a program is stored, and the program can be executed by a processor to implement the method according to any one of the first aspects.

[0061] In one embodiment, the present invention provides a staining method based on an existing staining machine, which can prevent the glass slides from drying out through a suitable staining process method, and the staining effect is stable and uniform.

[0062] In one embodiment, the present invention utilizes modern control software technology to achieve automation of the staining experiment, stable results of the staining experiment, no blank in the sample area, and no overstaining of hematoxylin, etc.

[0063] In one embodiment, the present invention effectively solves problems such as non-uniform and unstable results of the staining experiment, blank in the central area of the sample, overstaining of hematoxylin, and harsh requirements for reaction conditions.

[0064] Example 1

[0065] Figure 1 The following is a flowchart for controlling a staining machine to achieve cell staining, and the control method is as follows:

[0066] (1) Sequentially place the prepared sample slides into the slide rack and place them in the reaction water tank of the instrument. Set the sample volume in the instrument setting interface, select the repair experiment, and perform the antigen repair experiment.

[0067] (2) The instrument controls the heating component of the reaction water tank to raise the water tank temperature to the set temperature and maintain it for the set time, and then naturally cools down. After the cooling is completed, the reaction water tank is automatically opened.

[0068] (3) Take out the slide rack, forcefully shake off the residual repair solution on the slides, use a hydrophobic pen to supplement and draw hydrophobic circles, and drop cleaning solution in the sample area of the slides; after all the slides are processed, click the drainage on the instrument experiment setting interface to control the instrument to automatically drain the residual repair solution in the reaction water tank.

[0069] (4) After drainage is completed, sequentially place the slides into the reaction water tank. Set the sample volume in the instrument experiment setting interface, select the staining experiment step, and control the instrument to start the staining experiment.

[0070] (5) The specific process of the staining experiment is as follows:

[0071] A. Under the control of the internal control software of the instrument, the staining experiment starts. First, the instrument is controlled to perform pipeline filling and pipette cleaning, etc.

[0072] B. Control the multi-pipette arm of the instrument to add sufficient pure water to all the water tanks with samples, so that the slides in the water tanks are in a water bath state.

[0073] C. Control the multi-pipette arm, and use the stepping motor torque control technology to detect the positions of the slides, calculate the optimal cleaning height for each slide, so as to be able to completely clean the slides and not suck away the samples in the sample area of the slides.

[0074] D. The first round of cleaning: Control the multi-pipette arm of the instrument to perform PBS T cleaning on the slides with the cleaning solution according to the experimental set parameters.

[0075] E. Second-round cleaning: After the first-round cleaning is completed, control the single-pin pipetting arm to aspirate the DS-A reagent and dispense it onto each glass slide. After waiting for the reaction of the DS-A reagent to complete, perform the second-round PBST cleaning.

[0076] F. Third-round cleaning: After the second-round cleaning is completed, control the single-pin pipetting arm to aspirate the DS-B reagent and dispense it onto the glass slides. After waiting for the reaction of the DS-B reagent to complete, perform the third-round PBST cleaning.

[0077] G. Fourth-round cleaning: After the third-round cleaning is completed, control the single-pin pipetting arm to aspirate the DS-C reagent and dispense it onto the glass slides. After waiting for the reaction of the DS-C reagent to complete, perform the fourth-round PBST cleaning.

[0078] H. Prepare DAB reagent: During the interval of the fourth-round cleaning, control the single-pin pipetting arm to aspirate sufficient amounts of DS-D and DS-E reagents in a ratio of 1:33 and mix them into the DAB glass bottle to form the DAB reagent.

[0079] I. Fifth-round cleaning: After the fourth-round cleaning is completed, control the single-pin pipetting arm to aspirate the DAB reagent and dispense it onto the glass slides. After waiting for the reaction of the DAB reagent to complete, perform the fifth-round cleaning. Specifically, first clean the glass slides with pure water according to the set parameters. After the pure water cleaning is completed, continue to perform a PBST cleaning once.

[0080] J. During the interval of the fifth-round cleaning, control the single-pin pipetting arm to aspirate sufficient amounts of DS-F and DS-G reagents in a volume ratio of 1:50 and mix them into the Fast-Red glass bottle to prepare the Fast-Red reagent.

[0081] K. Sixth-round cleaning: After the fifth-round cleaning is completed, control the single-pin pipetting arm to aspirate the Fast-Red reagent and dispense it onto the glass slides. After waiting for the reaction of the Fast-Red reagent to complete, perform the sixth-round cleaning with pure water.

[0082] L. Seventh-round cleaning: After the sixth-round cleaning is completed, control the multi-pin pipetting arm to fill the hematoxylin liquid path with hematoxylin. Subsequently, control the multi-pin to dispense hematoxylin onto the glass slides. After dispensing in one reaction tank, wait for 1 second and immediately perform the seventh-round cleaning of the glass slides with pure water, and sequentially execute all tanks. After each tank is completed, perform the seventh-round cleaning of the corresponding other tanks with pure water.

[0083] M. Eighth-round cleaning: After the seventh-round cleaning is completed, control the multi-pin pipetting arm to fill the blueing solution (also known as the counterstain solution) pipeline with the blueing solution. Subsequently, control the multi-pin to dispense the blueing solution onto the glass slides. After waiting for the reaction of the blueing solution to complete, perform the eighth-round cleaning with pure water.

[0084] N. After the eighth round of cleaning is completed, control the instrument to empty the pipeline, reset all motion axes, open the sink cover, and pop up the experiment completion interface.

[0085] (6) After the experiment is completed, take out the glass slide and observe the cell staining under the microscope.

[0086] (7) The experiment throughput control method is as follows: Divide the experimental platform into 6 regions, and each region can be operated independently. The throughput can be freely selected from 1 to 30 glass slides, and the placement position of the glass slide experiment can be freely selected in the 6 regions.

[0087] (8) The cleaning method of the single-pin pipetting arm is as follows: When the single-pin pipetting arm is washed by flushing in pure water, slowly lift the pipetting needle to avoid water droplets hanging at the end of the pipetting needle, so as to contaminate the dilution reagent when sucking the reagent. Lift the pipetting needle until the tip of the single pin is higher than the pure water level.

[0088] (9) The liquid preparation method of DAB and Fast-Red reagents:

[0089] The ratio of DAB reagent DS-D:DS-E = 1:33 = d;

[0090] The ratio of Fast-Red reagent DS-F:DS-G = 1:50 = f;

[0091] The total amount of reagents S required for DAB and Fast-Red;

[0092] The volume of DS-D liquid = [S×d] + 1;

[0093] The volume of DS-E liquid = the volume of DS-D liquid × 33;

[0094] The volume of DS-F liquid = [S×f] + 1;

[0095] The volume of DS-G liquid = the volume of DS-D liquid × 55.

[0096] Through the above calculation method, it is possible to avoid problems caused by inaccurate reagent mixing ratios due to the inability to achieve a resolution accuracy of 0.1 level for the pipetting needle when sucking liquid volumes because the calculated values are of decimal types.

[0097] In this embodiment, the descriptions of each reagent are as follows:

[0098] Table 1

[0099]

[0100]

[0101] Table 2

[0102] Name Main Component DS-A Endogenous Peroxidase Blocker DS-B Antibody, Protein Stabilizer DS-C Antibody, Protein Stabilizer DS-D Diaminobenzidine DS-E Diaminobenzidine Diluent DS-F Fast red Chromogen DS-G Fast red Buffer

[0103] In one embodiment, the cell staining method provided by the present invention effectively improves the stability of cell staining effect.

[0104] In one embodiment, other cleaning methods can be used to avoid the problem that the samples in the sample area of the glass slide are sucked away during the cleaning process.

[0105] Those skilled in the art can understand that all or part of the functions of the various methods in the above embodiments can be implemented in a hardware manner or in a computer program manner. When all or part of the functions in the above embodiments are implemented in a computer program manner, the program can be stored in a computer-readable storage medium, and the storage medium can include: read-only memory, random access memory, magnetic disk, optical disk, hard disk, etc. The above functions are implemented by a computer executing the program. For example, the program is stored in the memory of the device, and when the processor executes the program in the memory, the above-mentioned all or part of the functions can be realized. In addition, when all or part of the functions in the above embodiments are implemented in a computer program manner, the program can also be stored in a storage medium such as a server, another computer, magnetic disk, optical disk, flash drive or mobile hard disk, and saved to the memory of the local device by downloading or copying, or the system of the local device is updated. When the processor executes the program in the memory, all or part of the functions in the above embodiments can be realized.

[0106] The above uses specific examples to elaborate on the present invention, which is only used to help understand the present invention and is not intended to limit the present invention. For those skilled in the technical field to which the present invention belongs, according to the idea of the present invention, several simple deductions, deformations or substitutions can also be made.

Claims

1. An immunohistochemical staining method, characterized in that, it includes: A cleaning height calculation step, placing the slide carrying the sample after antigen repair in a water tank, such that the lower surface of the slide is in the water bath and the sample area on the upper surface of the slide is not immersed in the water bath, using the stepping motor torque control technology to detect the position of the slide, calculating the optimal cleaning height of the slide, and setting parameters according to the optimal cleaning height; wherein, the optimal cleaning height refers to the height that can completely clean the slide without sucking away the sample in the sample area of the slide; A cleaning step, including controlling the pipetting arm, sucking reagents, dispensing them onto each slide, and after the reaction is completed, sucking the cleaning agent and cleaning the slide according to the set parameters to obtain the slide after immunohistochemical staining; Among them, in the cleaning step, it includes the first round of cleaning, the second round of cleaning, the third round of cleaning, the fourth round of cleaning, the fifth round of cleaning, the sixth round of cleaning, the seventh round of cleaning, and the eighth round of cleaning; for the second round and each subsequent round of cleaning, before each round of cleaning, control the pipetting arm, suck the required reagents to be added, dispense them onto the slide, and after the reagents react, clean the slide with the cleaning agent according to the set parameters; the reagent added before the second round of cleaning is the DS-A reagent, the reagent added before the third round of cleaning is the DS-B reagent, the reagent added before the fourth round of cleaning is the DS-C reagent, the reagent added before the fifth round of cleaning is the DAB reagent, the DAB reagent contains the DS-D reagent and the DS-E reagent, the reagent added before the sixth round of cleaning is the Fast-Red reagent, the Fast-Red reagent contains the DS-F reagent and the DS-G reagent, the reagent added before the seventh round of cleaning is the staining agent, and the reagent added before the eighth round of cleaning is the blueing solution.

2. The immunohistochemical staining method according to claim 1, characterized in that, the seventh round of cleaning includes: controlling the pipetting arm to suck the staining agent and dispensing it onto each slide, waiting for a certain time after the dispensing of the slides in one reaction water tank is completed, and then cleaning the slide with the cleaning agent according to the set parameters, and using the same method to dispense the staining agent to other water tanks and clean them.

3. The immunohistochemical staining method according to claim 2, characterized in that, the certain time is 0.5 - 2 s.

4. The immunohistochemical staining method according to claim 2, characterized in that, the staining agent includes hematoxylin.

5. The immunohistochemical staining method according to claim 2, characterized in that, the cleaning agent used in the seventh round of cleaning contains water.

6. The immunohistochemical staining method according to claim 1, characterized in that, according to the set parameters, controlling the pipetting arm to clean the slide to obtain the slide after immunohistochemical staining.

7. The immunohistochemical staining method according to claim 1, characterized in that, in the DAB reagent, by volume, the DS-D reagent: the DS-E reagent = 1:33, and this ratio is denoted as d; In the Fast-Red reagent, by volume, the DS-F reagent: the DS-G reagent = 1:50, and this ratio is denoted as f; The total amount of reagents required for the DAB reagent and the Fast-Red reagent is denoted as S; The liquid volume of the DS-D reagent = [S×d]+1; The liquid volume of the DS-E reagent = the liquid volume of the DS-D reagent × 33; The liquid volume of the DS-F reagent = [S×f]+1; The liquid volume of the DS-G reagent = the liquid volume of the DS-D reagent × 55.

8. The immunohistochemical staining method according to claim 1, characterized in that, The cleaning agent used in the first round of cleaning to the sixth round of cleaning contains PBST.

9. The immunohistochemical staining method according to claim 1, characterized in that, In the cleaning step, the cleaning agent used in the eighth round of cleaning contains water.

10. The immunohistochemical staining method according to claim 1, characterized in that, In the cleaning step, the experimental platform for placing the glass slides includes at least one area, and the glass slides in each area are stained separately.

11. The immunohistochemical staining method according to claim 10, characterized in that, In the cleaning step, the number of the areas is 1 to 10.

12. The immunohistochemical staining method according to claim 11, characterized in that, In the cleaning step, the number of the areas is 6.

13. The immunohistochemical staining method according to claim 1, characterized in that, In the cleaning step, when the pipetting arm is rinsed and cleaned in pure water, the pipetting needle is slowly lifted.

14. An immunohistochemical staining device, characterized in that, comprises: A cleaning height calculation module, which places the glass slide carrying the sample after antigen repair in a water tank so that the lower surface of the glass slide is in the water bath and the sample area on the upper surface of the glass slide is not immersed in the water bath, uses the stepping motor torque control technology to detect the position of the glass slide, calculates the optimal cleaning height of the glass slide, and sets parameters according to the optimal cleaning height; wherein, the optimal cleaning height refers to the height that can completely clean the glass slide and will not suck away the sample in the sample area of the glass slide; The cleaning module includes controlling the pipetting arm to aspirate reagents and dispense them onto each slide. After the reaction is completed, the cleaning agent is aspirated, and the slide is cleaned according to the set parameters to obtain the slide after immunohistochemical staining. Among them, in the cleaning step, it includes the first-round cleaning, the second-round cleaning, the third-round cleaning, the fourth-round cleaning, the fifth-round cleaning, the sixth-round cleaning, the seventh-round cleaning, and the eighth-round cleaning. For the second-round cleaning and each subsequent round of cleaning, before each round of cleaning, the pipetting arm is controlled to aspirate the required reagents and dispense them onto the slide. After the reaction of the reagents is completed, the slide is cleaned with the cleaning agent according to the set parameters. The reagent added before the second-round cleaning is the DS-A reagent, the reagent added before the third-round cleaning is the DS-B reagent, the reagent added before the fourth-round cleaning is the DS-C reagent, the reagent added before the fifth-round cleaning is the DAB reagent, and the DAB reagent contains the DS-D reagent and the DS-E reagent. The reagent added before the sixth-round cleaning is the Fast-Red reagent, and the Fast-Red reagent contains the DS-F reagent and the DS-G reagent. The reagent added before the seventh-round cleaning is the staining agent, and the reagent added before the eighth-round cleaning is the blueing solution.

15. An immunohistochemical staining device characterized in that it includes: a memory for storing programs; a processor for implementing the immunohistochemical staining method according to any one of claims 1 to 13 by executing the programs stored in the memory.

16. A computer-readable storage medium characterized in that a program is stored on the medium, and the program can be executed by a processor to implement the immunohistochemical staining method according to any one of claims 1 to 13.

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