A method for preparing an immunohistochemical antibody quality test chip

Abstract

The present invention relates to the technical field of test chip preparation, specifically a method for preparing an immunohistochemical antibody quality test chip, comprising the following steps: S1, chip microarray design: selecting a variety of positive and negative tissue specimens to design 15 points; designing a 5×3 point tissue array; S2, collecting relevant wax blocks: selecting the required tissue wax blocks, and marking representative points in the paraffin specimens according to HE sections. This application clearly explains the operating method and the technical issues that need to be paid attention to during the operation, as well as the time and temperature that need to be controlled, so as to fully guarantee the quality of the corresponding staff during the operation to ensure the test effect. At the same time, this method can effectively increase the number of test chips during the test, and can effectively meet the screening of high-specificity, high-affinity antibodies.

Description

Technical Field

[0001] The present invention relates to the technical field of test chip preparation, and in particular to a method for preparing an immunohistochemical antibody quality test chip. Background Art

[0002] With the advent of personalized diagnosis and treatment and the rapid development of precision medicine, immunohistochemistry has become an indispensable diagnostic aid for pathologists. Furthermore, with the increasing automation of the immunohistochemical staining process, the role of immunohistochemistry in pathological diagnosis is becoming increasingly widespread. Today, immunohistochemistry technology is not only used to determine the origin, classification, and differential diagnosis of tumors, but is also widely used to detect relevant indicators to determine prognosis and guide personalized clinical treatment. Therefore, its accuracy and reliability are crucial. While immunohistochemistry has been used in clinical pathology for over 40 years, standardization of quality control still requires further improvement. Numerous factors can affect the immunohistochemical staining process to varying degrees. Over 70% of the accuracy and reliability of immunohistochemistry is determined by the quality of antibody reagents. Therefore, quality control by antibody manufacturers is a crucial factor in ensuring the accuracy and reliability of immunohistochemical results. Currently, the quality of antibodies produced by domestic immunohistochemical manufacturers varies greatly. To improve the reliability of staining results and ensure the effectiveness of immunohistochemical staining antibodies and related reagents, we have developed this tissue microarray to validate antibodies.

[0003] Currently, immunohistochemistry antibody manufacturers face the following problems in antibody quality control: low standardization and high uncertainty in immunohistochemistry; a lot of effort is required to prepare the positive control tissues required for management control; quality control tissues required for some targets are difficult to obtain; tissue differences add more uncontrollable processes to the quality control process; ethical issues make the quality control system difficult to translate. Currently, immunohistochemistry antibody manufacturers conduct quality inspections on the antibodies they produce by verifying them on a single tissue section. The advantage of this verification method is that the specimen area used for testing is large, and the positive area is generally large. However, high-quality pathological diagnostic antibodies require verification with multiple tissue specimens, and negative controls must be set up. A single tissue section cannot meet the screening requirements for high-specificity and high-affinity antibodies, so improvements are needed. Summary of the Invention

[0004] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a method for preparing an immunohistochemical antibody quality test chip.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] A method for preparing an immunohistochemical antibody quality test chip comprises the following steps:

[0007] S1. Chip microarray design: Select a variety of positive and negative tissue specimens and design 15 spots; design a 5×3 spot tissue array;

[0008] S2. Collect relevant wax blocks: Select the required tissue wax blocks and mark representative points in the paraffin specimen according to the HE section;

[0009] S3. Preparation of TMA receptor wax blocks: Mix 97.5 g of paraffin wax and 2.5 g of beeswax to create a blank wax block measuring 36 mm long, 26 mm wide, and 17 mm high. Within the 20 mm x 16 mm area of ​​this wax block, design a 5 x 3 point tissue array. Leave 0.5 cm to 0.7 cm of space around the tissue and punch holes using a tissue analyzer to create a TMA wax block.

[0010] S4. Use a fine needle to punch holes in the recipient wax block on a tissue chip production machine. The hole diameter is designed to be 5 mm.

[0011] S5. Punch a hole at the marked location on the donor wax block to collect a tissue core; the hole diameter is also 5 mm.

[0012] S6. Transfer the tissue cores to the wells of the receptor module, with the spacing between each tissue core preferably being 0.5 mm;

[0013] S7. Place the constructed TMA chip wax block in a suitable plastic box and secure it tightly to prevent displacement. Place it in a 55°C incubator for approximately 10 minutes. Before the wax completely dissolves (half-melted), remove it and cool it at room temperature to allow the wax of the receptor module and the newly inserted small cylindrical tissue to dissolve into one. Remove the wax block and store it in a 4°C refrigerator until needed.

[0014] S8. Before slicing, the wax block needs to be pre-cooled at 4℃ for about 4 hours, then clamped on the microtome for correction until all the tissues are complete; stick a -20℃ pre-cooled ice bag on the wax block for about 5-10 minutes, quickly slice about 30-50 slices in succession, and then freeze the tissue block with an ice bag. It can also be done directly in the freezing microtome until all the tissues are cut; float 4μm serial sections in cold water and let them unfold naturally. Transfer the sections to 45℃ warm water in sequence and unfold them for about 2 minutes. Stick them on a slide soaked in APES section adhesive to dry, bake the sections at 60℃ for about 3 minutes, continue baking at 58℃ for 18 hours, and store them at -20℃ for later use.

[0015] Compared with the existing technology, this application clearly explains the operation method, the technical issues that need to be paid attention to during operation, and the time and temperature that need to be controlled, so as to fully ensure the quality of the operation of the relevant staff to ensure the test effect. At the same time, this method can effectively increase the number of test chips during the test, and can effectively meet the screening of high-specificity and high-affinity antibodies.

[0016] Preferably, three points located on the same side of the 15 points in S1 form a group, and the five groups of points use tonsils, liver, appendix, kidneys, and pancreas as templates respectively.

[0017] Furthermore, a control group can be effectively formed.

[0018] Preferably, the plastic box in S7 includes a shell, a partition member is fixed in the shell, an adjustment mechanism is provided at the lower end of the shell, a first gear member is provided on the adjustment mechanism, four first straight racks are meshed on the first gear member from top to bottom, and two adjacent first straight racks are arranged at right angles, the first straight racks are slidably installed on the lower end of the partition member, an adaptation mechanism is provided on the first straight rack, a synchronization rod member is provided on the adaptation mechanism, four openings are equidistantly provided on the partition member, four synchronization rod members are respectively slidably installed in the four openings, and a splint member is fixed on the upper end of the four synchronization rod members.

[0019] Furthermore, the movement of the clamping plate can be controlled by the functions of the regulating mechanism and the adapting mechanism, and the TMA chip wax block can be effectively clamped by the movement of the clamping plate.

[0020] Preferably, the adjustment mechanism includes a mounting bracket installed at the bottom of the shell, a second spur rack is slidably mounted on one side of the mounting bracket, a second gear member is meshed on the second spur rack, a vertical shaft member is fixedly sleeved inside the second gear member, the lower end of the vertical shaft member is rotatably connected to the bottom center of the shell, the upper end of the vertical shaft member is fixed to the lower end of the first gear member, a screw member is rotatably sleeved on one side of the lower end of the shell, and one side of the second spur rack is screwed on the screw member.

[0021] Furthermore, it is convenient for workers to quickly rotate the screw member so as to move the splint member through the cooperation of corresponding parts.

[0022] Preferably, the adaptation mechanism includes a fixed plate fixed on one side of the first straight rack, a sliding rod is slidably installed on the fixed plate, one end of the sliding rod is fixedly connected to the synchronization rod, a spring is sleeved on the sliding rod, and both ends of the spring are respectively fixed on the fixed plate and the sliding rod.

[0023] Furthermore, through the action of the spring member and the slide rod member, it can adapt to TMA chip wax blocks of various specifications and better clamp them.

[0024] Preferably, the diameter of the first gear member is greater than the diameter of the second gear member.

[0025] Furthermore, due to the diameter difference between the first gear member and the second gear member, the rotational circumference of the first gear member, that is, the number of teeth, is greater, thereby increasing the moving speed of the first spur rack.

[0026] Preferably, the mixture of 97.5 grams of lycra paraffin wax and 2.5 grams of beeswax in S3 is prepared by a mixing mechanism, the mixing mechanism includes a carrier frame, both sides of the carrier frame are rotatably sleeved with rotating shafts, a regulating mechanism is installed in the carrier frame, the regulating mechanism is connected to one of the rotating shafts, a hopper member is fixed between the two rotating shafts, the lower end of the hopper member is provided with a power mechanism, the power mechanism is provided with a lifting rod and a vertical shaft member, the vertical shaft member is rotatably sleeved on the hopper member, the upper end of the lifting rod is fixed with a cross bar member, one end of the cross bar member is rotatably sleeved with a sleeve member, the sleeve member is slidably mounted on the vertical shaft member, a stirring member is installed on the sleeve member, a mounting plate member is fixed to one side of the lower end of the vertical shaft member, a scraper member is installed on the mounting plate member, and the scraper member and the inner wall of the hopper member are in conflict.

[0027] Furthermore, the automatic mixing effect of Lycra paraffin and beeswax can be effectively guaranteed.

[0028] Preferably, the regulating mechanism includes a hydraulic cylinder component fixed to the bottom of the support frame, a third spur rack is fixed to the end of the piston rod of the hydraulic cylinder component, a rotating gear component is fixed on one of the rotating shaft components, the third spur rack and the rotating gear component are meshed, and the third spur rack is slidably mounted on one end side wall of the support frame.

[0029] Furthermore, by pushing and pulling the third spur rack to lift and lower it through the hydraulic cylinder, the rotating gear part can drive one of the rotating shaft parts to rotate forward and reverse, so as to quickly swing the hopper part and discharge the wax liquid therein.

[0030] Preferably, the power mechanism includes a connecting frame installed on one side of the lower end of the hopper member, a motor assembly is installed on one side of the connecting frame, the output shaft of the motor assembly and the vertical shaft member are coaxially arranged, the output shaft of the motor assembly and the lower end of the vertical shaft member are jointly fixed with a U-shaped rod, a push rod member is rotatably sleeved on the U-shaped rod, one end of the push rod member is rotatably connected to an oblique rod member, the lower end of the oblique rod member is rotatably connected to the lower end of the lifting rod, and the lifting rod is slidably installed on one side of the hopper member.

[0031] Furthermore, the motor assembly can provide power for scraping the inner wall of the hopper member and mixing the materials in the hopper member.

[0032] Preferably, an electric heater is installed on the hopper member.

[0033] Furthermore, the electric heater can increase the temperature in the hopper, which helps to form the wax into wax liquid, making it easier to prepare it into a suitable form as needed.

[0034] The beneficial effects of the present invention are:

[0035] 1. By clarifying the method for preparing the test chip, subsequent staff can accurately follow the steps, time and temperature, fully ensuring the efficiency and quality of chip production. At the same time, multiple test chips can be effectively formed for effective control, meeting the screening of high-specificity and high-affinity antibodies, and improving the effect and quality of the test;

[0036] 2. By clarifying the items used in preparation, the preparation effect can be effectively guaranteed and the preparation efficiency can be fully improved;

[0037] 3. By stirring and heating, the mixing effect of Lycra paraffin and beeswax can be fully improved, and at the same time, the mixed wax liquid can be quickly discharged to facilitate its molding. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 It is a block diagram of the steps of the present invention;

[0039] Figure 2 Schematic diagram of sample points of the present invention;

[0040] Figure 3 is a cross-sectional view of the carrier box of the present invention;

[0041] Figure 4 The present invention is attached Figure 3 A magnified view of point A;

[0042] Figure 5 The present invention is attached Figure 3 Enlarged view of point B;

[0043] Figure 6 It is a structural diagram of the hybrid mechanism of the present invention;

[0044] Figure 7 The present invention is attached Figure 6 Enlarged view of point C;

[0045] In the figure: 1 shell, 2 partition member, 3 opening, 4 first straight rack, 5 second straight rack, 6 screw member, 7 first gear member, 8 clamping plate member, 9 slide rod member, 10 synchronization rod member, 11 spring member, 12 fixed plate, 13 vertical shaft member, 14 second gear member, 15 mounting frame, 16 cross bar member, 17 lifting rod, 18 bearing frame, 19 rotating shaft member, 20 push rod member, 21 oblique rod member, 22 rotating gear member, 23 third straight rack, 24 hydraulic cylinder member, 25 motor assembly, 26 connecting frame, 27 mounting plate member, 28 scraper member, 29 vertical shaft member, 30 hopper member, 31 U-shaped rod, 32 stirring member, 33 sleeve member, 34 electric heater. DETAILED DESCRIPTION

[0046] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0047] Reference Figure 1-2 A method for preparing an immunohistochemical antibody quality test chip comprises the following steps:

[0048] S1. Chip microarray design: Select a variety of positive and negative tissue specimens and design 15 spots. Design a 5×3 spot tissue array, with 3 spots on the same side of the 15 spots forming a group. Five groups of spots use tonsils, liver, appendix, kidney, and pancreas as templates. This can effectively expand the sample range and enable testing on multiple tissue sections, effectively meeting the screening requirements for highly specific and high-affinity antibodies.

[0049] S2. Collect relevant wax blocks: Select the required tissue wax blocks and mark representative points in the paraffin specimen according to the HE section;

[0050] S3. Preparation of TMA receptor wax blocks: Mix 97.5 g of paraffin wax and 2.5 g of beeswax to create a blank wax block measuring 36 mm long, 26 mm wide, and 17 mm high. Within the 20 mm x 16 mm area of ​​this wax block, design a 5 x 3 point tissue array. Leave 0.5 cm to 0.7 cm of space around the tissue and punch holes using a tissue analyzer to create a TMA wax block.

[0051] S4. Use a fine needle to punch holes in the recipient wax block on a tissue chip production machine. The hole diameter is designed to be 5 mm.

[0052] S5. Punch a hole at the marked location on the donor wax block to collect a tissue core; the hole diameter is also 5 mm.

[0053] S6. Transfer the tissue cores to the wells of the receptor module, with the spacing between each tissue core preferably being 0.5 mm;

[0054] S7. Place the constructed TMA chip wax block in a suitable plastic box and secure it tightly to prevent displacement. Place it in a 55°C incubator for approximately 10 minutes. Before the wax completely dissolves (half-melted), remove it and cool it at room temperature to allow the wax of the receptor module and the newly inserted small cylindrical tissue to dissolve into one. Remove the wax block and store it in a 4°C refrigerator until needed.

[0055] S8. Before slicing, the wax block needs to be pre-cooled at 4℃ for about 4 hours, then clamped on the microtome for correction until all the tissues are complete; stick a -20℃ pre-cooled ice bag on the wax block for about 5-10 minutes, quickly slice about 30-50 slices in succession, and then freeze the tissue block with an ice bag. It can also be done directly in the freezing microtome until all the tissues are cut; float 4μm serial sections in cold water and let them unfold naturally. Transfer the sections to 45℃ warm water in sequence and unfold them for about 2 minutes. Stick them on a slide soaked in APES section adhesive to dry, bake the sections at 60℃ for about 3 minutes, continue baking at 58℃ for 18 hours, and store them at -20℃ for later use.

[0056] Reference Figure 3-5 The plastic box in S7 includes a shell 1, a partition member 2 is fixed in the shell 1, an adjustment mechanism is provided at the lower end of the shell 1, and a first gear member 7 is provided on the adjustment mechanism. The adjustment mechanism includes a mounting frame 15 installed at the bottom of the shell 1, a second spur rack 5 is slidably mounted on one side of the mounting frame 15, and a second gear member 14 is engaged with the second spur rack 5. A vertical shaft member 13 is fixedly sleeved in the second gear member 14, and the lower end of the vertical shaft member 13 is rotatably connected to the bottom center of the shell 1. The upper end of the vertical shaft member 13 is fixed to the lower end of the first gear member 7. The diameter of the first gear member 7 is larger than that of the second gear member 14. Diameter. By changing the diameter, when the first gear member 7 and the second gear member 14 rotate at the same angle, the number of teeth of the first gear member 7 rotates more than that of the second gear member 14, which helps to achieve rapid movement of the first spur rack 4. A screw member 6 is rotatably sleeved on one side of the lower end of the shell 1, and one side of the second spur rack 5 is screwed on the screw member 6. By rotating the screw member 6, the second spur rack 5 can be moved smoothly. When the second spur rack 5 moves smoothly, the second gear member 14 can be rotated smoothly to drive the first gear member 7 to rotate, thereby providing power for the movement of the splint member 8.

[0057] Reference Figure 3-5, four first straight racks 4 are meshed in sequence on the first gear member 7 from top to bottom, and the two adjacent first straight racks 4 are arranged at right angles. The four first straight racks 4 can effectively correspond to the four sides of the rectangular structure, which helps to clamp the rectangular structure by clamping the four sides. The first straight rack 4 is slidably installed at the lower end of the partition member 2, which can ensure the stability of the movement of the first straight rack 4. An adaptation mechanism is provided on the first straight rack 4, and a synchronization rod 10 is provided on the adaptation mechanism. Four openings 3 are equally spaced on the partition member 2. The four synchronization rods 10 are respectively slidably installed in the four openings 3. The upper ends of the four synchronization rods 10 are fixed with a clamping plate 8. The adaptation mechanism includes a fixed plate 12 fixed to one side of the first straight rack 4, and a sliding rod 9 is slidably installed on the fixed plate 12. One end of the sliding rod 9 and the synchronization The rod 10 is fixedly connected, and a spring part 11 is sleeved on the sliding rod 9. The two ends of the spring part 11 are respectively fixed on the fixed plate 12 and the sliding rod 9. Through the action of the spring part 11, the splint part 8 can stop moving after it hits the material. If there is a deviation in other positions, when the splint part 8 hits one side of the material, the other external splint parts 8 have not yet hit the material. At this time, the material can be pushed to move, so that the two splint parts 8 set at a right angle can hit the material. If the other two splint parts 8 have not yet hit the material, the spring parts 11 at the lower ends of the two splint parts 8 that have hit can be stretched so that the other two splint parts 8 move and hit the material, which can effectively ensure the firmness of the clamping. The spring part 11 can be telescopic to adapt to the movement of the first straight rack 4, and can effectively clamp the four sides of the material.

[0058] Reference Figure 6-7 The mixture of 97.5 grams of lycra paraffin wax and 2.5 grams of beeswax in S3 is prepared by a mixing mechanism. The mixing mechanism includes a carrier 18. Both sides of the carrier 18 are rotatably sleeved with rotating shafts 19. A regulating mechanism is installed in the carrier 18. The regulating mechanism is connected to one of the rotating shafts 19. A hopper 30 is fixed between the two rotating shafts 19. The regulating mechanism can adjust one of the rotating shafts 19 to rotate the rod, which can drive the hopper 30 to flip it. When the hopper 30 flips, it is convenient to add materials or discharge the melted wax liquid so that the wax liquid can be formed.

[0059] Reference Figure 6The regulating mechanism includes a hydraulic cylinder 24 fixed to the bottom of the carrier 18. A third spur rack 23 is fixed to the end of the piston rod of the hydraulic cylinder 24. A rotating gear 22 is fixed to one of the rotating shafts 19. The third spur rack 23 and the rotating gear 22 are meshed. The third spur rack 23 is slidably mounted on one end side wall of the carrier 18. During actual preparation, the staff can make the hydraulic cylinder 24 push the third spur rack 23 to move up and down. When the third spur rack 23 moves up and down, it will drive the rotating gear 22 meshed with it to rotate. The rotating gear 22 can drive the rotating shaft 19 fixed to it to rotate. The rotation of the rotating shaft 19 causes the hopper 30 to flip, so that the molten wax liquid inside can be quickly discharged and the wax liquid can be formed. During preparation, a protruding mouth-shaped structure can be provided on the hopper 30 to better facilitate the stable outflow of materials.

[0060] Reference Figure 6 An electric heater 34 is installed on the hopper part 30. The electric heater 34 can increase the temperature inside the hopper part 30, which helps to melt the lycra paraffin and beeswax and mix them better. A power mechanism is provided at the lower end of the hopper part 30. The power mechanism is provided with a lifting rod 17 and a vertical shaft part 29. The vertical shaft part 29 is rotatably sleeved on the hopper part 30. A cross bar 16 is fixed to the upper end of the lifting rod 17. One end of the cross bar 16 is rotatably sleeved with a sleeve part 33. The sleeve part 33 is slidably installed on the vertical shaft part 29. A stirring member 32 is installed on the sleeve part 33. A mounting plate is fixed to one side of the lower end of the vertical shaft part 29 Part 27, a scraper part 28 is installed on the mounting plate part 27, and the scraper part 28 conflicts with the inner wall of the hopper part 30. The power mechanism includes a connecting frame 26 installed on one side of the lower end of the hopper part 30, and a motor assembly 25 is installed on one side of the connecting frame 26. The output shaft of the motor assembly 25 and the vertical shaft part 29 are coaxially arranged, and the output shaft of the motor assembly 25 and the lower end of the vertical shaft part 29 are jointly fixed with a U-shaped rod 31. The coaxial arrangement of the output shaft of the motor assembly 25 and the vertical shaft part 29 can fully ensure the stable transmission of power through the U-shaped rod 31, and ensure that the vertical shaft part 29 can keep up with the synchronous rotation of the output shaft of the motor assembly 25.

[0061] During use, a worker can start the motor assembly 25 to rotate it. The rotation of the motor assembly 25 can cause the U-shaped rod 31 to drive the vertical shaft 29 to rotate, and the vertical shaft 29 can drive the mounting plate 27 and the scraper 28 to rotate. The scraper 28 can clean the inner wall of the hopper 30, effectively cleaning the material in the hopper 30, ensuring the cleanliness of the hopper 30 and facilitating the discharge of the material.

[0062] The U-shaped rod 31 is rotatably sleeved with a push rod member 20, and one end of the push rod member 20 is universally connected to an oblique rod member 21. The lower end of the oblique rod member 21 is rotatably connected to the lower end of the lifting rod 17. The lifting rod 17 is slidably installed on one side of the hopper member 30, which can ensure the smooth lifting and lowering of the lifting rod 17. The push rod member 20 can be driven to operate through the action of the U-shaped rod 31. At the same time, the oblique rod member 21 is swung through the corresponding universal connection member, which can effectively provide power for the lifting and lowering of the lifting rod 17. When the lifting rod 17 is lifted or lowered, it can drive the cross rod member 16 to lift the sleeve member 33. At the same time, the sleeve member 33 rotates with the vertical shaft member 29, and the sleeve member 33 will drive the stirring member 32 to rotate, so that the stirring member 32 can stir and mix the Lycra paraffin and beeswax.

[0063] In the present invention, 15 points are designed from a variety of positive and negative tissue specimens; a 5×3 point tissue array is designed. The required tissue wax blocks are selected, and representative points in the paraffin specimens are marked according to HE sections. 97.5 grams of paraffin wax and 2.5 grams of beeswax are mixed to form a blank wax block measuring 36 mm long, 26 mm wide, and 17 mm high. A 5×3 point tissue array is designed within the 20 mm × 16 mm area of ​​this wax block. A 0.5 cm to 0.7 cm space is reserved around the tissue. A tissue analyzer is used to punch holes to create a TMA wax block. A fine needle is used to punch holes in the recipient wax block on a tissue chip making machine, with a designed hole diameter of 5 mm. Similarly, holes are punched at the corresponding locations marked on the donor wax block to collect tissue cores. The hole diameter is also 5 mm. The tissue cores are transferred to the holes of the recipient module, with a spacing of 0.5 mm between each tissue core being preferred. The constructed TMA chip wax block is placed in a suitable plastic box and tightly secured to prevent displacement. Place in a 55°C incubator for about 10 minutes. Before the wax is completely dissolved (half-melted), take it out and cool it at room temperature to allow the wax of the receptor module to dissolve into the newly inserted small cylindrical tissue. Remove the wax block and store it in a 4°C refrigerator for later use. Before slicing, the wax block needs to be pre-cooled at 4°C for about 4 hours, then clamped on the microtome for correction until all the tissue is complete. Apply a -20°C pre-cooled ice pack to the wax block for about 5-10 minutes, quickly slice 30-50 slices continuously, then freeze the tissue block with an ice pack, or directly in the freezing microtome until the tissue is cut. Float 4μm serial sections in cold water and let them unfold naturally. Transfer the sections to 45°C warm water in sequence and spread them for about 2 minutes. Attach them to a slide soaked in APES sectioning adhesive to dry. Bake the sections at 60°C for about 3 minutes, continue baking at 58°C for 18 hours, and store at -20°C for later use.

[0064] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solution and inventive concept of the present invention, should be covered by the scope of protection of the present invention.

Claims

1. A method for preparing an immunohistochemical antibody quality test chip, characterized in that: The following steps are involved: S1. Microarray design: Select various positive and negative tissue specimens and design 15 spots; design a 5×3 spot tissue array; 3 spots on the same side of the 15 spots form a group, and five groups of spots use tonsils, liver, appendix, kidney, and pancreas as templates respectively; S2. Collect relevant wax blocks: Select the required tissue wax blocks and mark representative points in the paraffin specimen according to the HE section; S3. Preparation of TMA receptor wax block: Mix 97.5 g of paraffin wax and 2.5 g of beeswax to make a blank wax block of 36 mm long, 26 mm wide, and 17 mm high. Design a 5 × 3 dot tissue array within the 20 mm × 16 mm area of ​​the wax block. Leave 0.5cm-0.7cm space around the tissue and use a tissue analyzer to punch holes to make TMA wax blocks; 97.5g of Klein wax + 2.5g of beeswax are mixed and prepared by a mixing mechanism; The mixing mechanism comprises a carrier (18), both sides of the carrier (18) are rotatably sleeved with rotating shafts (19), a regulating mechanism is installed in the carrier (18), the regulating mechanism is connected to one of the rotating shafts (19), a hopper (30) is fixed between the two rotating shafts (19), a power mechanism is provided at the lower end of the hopper (30), a lifting rod (17) and a vertical shaft (29) are provided on the power mechanism, and the vertical shaft (29) is rotatably sleeved on the hopper ( 30), a cross bar (16) is fixed to the upper end of the lifting rod (17), a sleeve (33) is rotatably sleeved on one end of the cross bar (16), the sleeve (33) is slidably mounted on the vertical shaft (29), a stirring member (32) is mounted on the sleeve (33), a mounting plate (27) is fixed to one side of the lower end of the vertical shaft (29), a scraper (28) is mounted on the mounting plate (27), and the scraper (28) is in contact with the inner wall of the hopper (30); The regulating mechanism includes a hydraulic cylinder member (24) fixed to the bottom of the carrier (18), a third spur rack (23) is fixed to the end of the piston rod of the hydraulic cylinder member (24), a rotating gear member (22) is fixed to one of the rotating shaft members (19), the third spur rack (23) and the rotating gear member (22) are meshed, and the third spur rack (23) is slidably mounted on a side wall at one end of the carrier (18); The power mechanism includes a connecting frame (26) installed on one side of the lower end of the hopper member (30), a motor assembly (25) is installed on one side of the connecting frame (26), an output shaft of the motor assembly (25) and a vertical shaft member (29) are coaxially arranged, and a U-shaped rod (31) is fixed to the lower ends of the output shaft of the motor assembly (25) and the vertical shaft member (29), a push rod member (20) is rotatably sleeved on the U-shaped rod (31), one end of the push rod member (20) is rotatably connected to an inclined rod member (21), the lower end of the inclined rod member (21) is rotatably connected to the lower end of a lifting rod (17), and the lifting rod (17) is slidably installed on one side of the hopper member (30); An electric heater (34) is installed on the hopper member (30); S4. Use a fine needle to punch holes in the recipient wax block on a tissue chip production machine. The hole diameter is designed to be 5 mm. S5. Punch a hole at the marked location on the donor wax block to collect a tissue core; the hole diameter is also 5 mm. S6. Transfer the tissue cores to the wells of the receptor module, with the spacing between each tissue core preferably being 0.5 mm; S7. Place the constructed TMA chip wax block in a suitable plastic box and secure it tightly to prevent displacement. Place it in a 55°C incubator for 10 minutes. When the wax is semi-melted, remove it and cool it at room temperature to allow the wax of the receptor module and the newly inserted small cylindrical tissue to dissolve into one. Remove the wax block and store it in a 4°C refrigerator until needed. The plastic box comprises a shell (1), a partition member (2) is fixed in the shell (1), an adjustment mechanism is provided at the lower end of the shell (1), a first gear member (7) is provided on the adjustment mechanism, four first straight racks (4) are meshed with the first gear member (7) in sequence from top to bottom, two adjacent first straight racks (4) are arranged at right angles, the first straight racks (4) are slidably mounted on the lower end of the partition member (2), an adaptation mechanism is provided on the first straight rack (4), a synchronization rod member (10) is provided on the adaptation mechanism, four openings (3) are provided on the partition member (2) at equal intervals, four synchronization rod members (10) are respectively slidably mounted in the four openings (3), and a clamping plate member (8) is fixed to the upper end of each of the four synchronization rod members (10); The adjustment mechanism comprises a mounting frame (15) mounted on the bottom of the housing (1), a second spur rack (5) being slidably mounted on one side of the mounting frame (15), a second gear member (14) being meshed with the second spur rack (5), a vertical shaft member (13) being fixedly sleeved inside the second gear member (14), the lower end of the vertical shaft member (13) being rotatably connected to the bottom center of the housing (1), the upper end of the vertical shaft member (13) being fixed to the lower end of the first gear member (7), a screw member (6) being rotatably sleeved on one side of the lower end of the housing (1), and one side of the second spur rack (5) being screwed onto the screw member (6); The adaption mechanism includes a fixed plate (12) fixed to one side of the first straight rack (4), a slide bar (9) is slidably mounted on the fixed plate (12), one end of the slide bar (9) is fixedly connected to the synchronization bar (10), a spring member (11) is sleeved on the slide bar (9), and both ends of the spring member (11) are fixed to the fixed plate (12) and the slide bar (9), respectively; The diameter of the first gear member (7) is greater than the diameter of the second gear member (14); S8. Before sectioning, the wax block needs to be pre-cooled at 4°C for 4 hours, then clamped on the microtome and trimmed until all tissues are intact; Apply a -20°C pre-cooled ice pack to the wax block for 5-10 minutes, quickly slice 30-50 slices in succession, and then freeze the tissue block with an ice pack; float 4μm serial sections in cold water and allow them to unfold naturally. Transfer the sections sequentially to 45°C warm water and unfold them for 2 minutes. Attach them to a slide soaked in APES section adhesive and let them dry. Bake the sections at 60°C for 3 minutes, continue baking at 58°C for 18 hours, and store at -20°C for later use.

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