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CN-121977912-A - Repairing liquid suitable for full-automatic immunohistochemical staining instrument and application method thereof

CN121977912ACN 121977912 ACN121977912 ACN 121977912ACN-121977912-A

Abstract

The invention provides a repairing liquid suitable for a full-automatic immunohistochemical staining instrument and a use method thereof, and particularly relates to the technical field of immunodetection, wherein the repairing liquid comprises, by mass, 0.1-1% of Tris, 0.01-0.1% of EDTA, 0.5-1% of glucan-20, 15-25% of ethylene glycol, 0.01-0.1% of Tween-20 and the balance of pure water, the temperature is set to be 60-100 ℃ in an upper program, the adding amount of the repairing liquid is 150-250 mu L, the reaction time is 30-50min, a tissue slice sample is used, the instrument runs automatically, the repairing liquid is not easy to volatilize and is environment-friendly, the storage condition is simple, meanwhile, the cleaning is easy, the strength and the background of the matched instrument are ideal, and the requirements of high-throughput and high-quality tabletting in hospitals can be met.

Inventors

  • LV PING
  • JIANG LINYE
  • WU XUYANG

Assignees

  • 卡秋(江苏)生物科技有限公司

Dates

Publication Date
20260505
Application Date
20251222

Claims (8)

  1. 1. A repairing liquid for the full-automatic immunohistochemical staining instrument is prepared from Tris, EDTA, dextran-20, ethylene glycol, tween-20 and purified water, and has pH of 9.0.
  2. 2. The repairing liquid for the full-automatic immunohistochemical staining instrument according to claim 1 is characterized by comprising, by mass, 0.1-1% of Tris, 0.01-0.1% of EDTA, 0.5-1% of glucan-20, 15-25% of ethylene glycol, 0.01-0.1% of tween-20 and the balance of pure water.
  3. 3. The repairing liquid for the full-automatic immunohistochemical staining instrument according to claim 1 is characterized by comprising the following substances by mass percent of 0.5% of Tris, 0.05% of EDTA, 0.75% of glucan-20, 20% of glycol and 0.05% of tween-20, and the balance of pure water.
  4. 4. The repairing liquid for the full-automatic immunohistochemical staining instrument according to claim 1 is characterized by comprising the following steps of setting the temperature to be 60-100 ℃ in an upper computer program, adding 150-250 mu L of the repairing liquid, reacting for 30-50min, and automatically operating the instrument by using a tissue section sample.
  5. 5. The method of claim 4, wherein the tissue slice is a paraffin-embedded tonsil tissue slice sample (CK 5/6) or a paraffin-embedded tonsil tissue slice sample (Ki 67) or a colon cancer tissue slice sample (Villin).
  6. 6. The method for using the repairing liquid suitable for the full-automatic immunohistochemical staining apparatus according to claim 4, wherein the method comprises the following steps: the temperature was set at 80 ℃ during the upper program.
  7. 7. The method of using a prosthetic liquid for a fully automatic immunohistochemical staining apparatus according to claim 4 wherein the prosthetic liquid is added in an amount of 200. Mu.L.
  8. 8. The method of using a prosthetic liquid for a fully automatic immunohistochemical staining apparatus according to claim 4 wherein the reaction time is 40min.

Description

Repairing liquid suitable for full-automatic immunohistochemical staining instrument and application method thereof Technical Field The invention belongs to the technical field of immunodetection, and particularly relates to a repair liquid suitable for a full-automatic immunohistochemical staining instrument. Background The early diagnosis of benign and malignant tumors in clinic is mainly judged by the staining result of pathological tissue sections. The doctor can make a slice or smear on the tissue suspected to be diseased, observe the tissue under a microscope after dyeing, think about the cause, mechanism and occurrence process of the diseased, and finally make pathological diagnosis. In general, the pathological tissue tableting process comprises material drawing, fixing, dehydration, transparency, wax dipping, embedding, slicing, dewaxing, dyeing, re-dehydration, re-transparency and sealing. In the above steps, the repair process plays a crucial role, and the selection of an appropriate repair liquid has a clear effect on the final result. Antigen retrieval refers to recovery of antigen recognizability and immunoreactivity in a tissue sample by heating or other methods. If this step is not done, the antigen may remain in an inactivated state, resulting in an inability of the antibody to specifically bind thereto. Antigen retrieval can increase the expression level of the target antigen in immunohistochemical experiments and enhance the signal intensity. Insufficient or inadequately optimized repair steps may result in a weakening of the signal, making the target antigen difficult to detect or reducing the signal intensity. In addition, existing antigen retrieval methods may cause damage or destruction to the tissue sample. Excessive heating or digestion time may result in destruction of tissue structures, rendering the cell and tissue morphology unrecoverable. Disclosure of Invention The invention aims to provide the repairing liquid suitable for the full-automatic immunohistochemical staining instrument, which is not easy to volatilize, is environment-friendly, has simple storage condition, is easy to clean, has ideal tabletting strength and background when matched with the instrument, and can meet the requirements of high throughput and high quality in hospitals. The invention provides a technical scheme that a repair liquid suitable for a full-automatic immunohistochemical staining instrument consists of Tris, EDTA, dextran-20, glycol, tween-20 and pure water, wherein the pH is 9.0. Preferably, the composition comprises (by weight) Tris 0.1-1%, EDTA 0.01-0.1%, and EDTA 0.5-1% Dextran-20 of (a), ethylene glycol of 15-25%, tween-20 of 0.01-0.1%, and pure water in balance. Preferably, the material comprises, by mass, 0.5% Tris, 0.05% EDTA, 0.75% dextran-20, 20% ethylene glycol and 0.05% Tween-20, and the balance pure water. A repairing liquid for full-automatic immunohistochemical staining instrument is prepared through setting the temp at 60-100 deg.C in upper program, adding 150-250 μL of repairing liquid, reacting for 30-50min, and automatic operation. Preferably, the tissue section is one of a paraffin-embedded tonsil tissue section sample (CK 5/6) or a paraffin-embedded tonsil tissue section sample (Ki 67) or an embedded colon cancer tissue section sample (Villin). Preferably, the temperature is set at 80 ℃ during the power-on procedure. Preferably, the repair liquid is added in an amount of 200. Mu.L. Preferably, the reaction time is 40min. The invention has the beneficial effects that: 1. the repairing liquid can effectively remove cross-linking among proteins caused by aldehyde fixing reagents, and fully expose epitope in samples such as paraffin sections, thereby greatly improving the immunostaining effect; 2. The repairing liquid provided by the invention has a full display effect on intracellular antigens, can improve the exposure degree of antigens which are difficult to detect, has the functions of moisturizing and easy adjustment, can better expose and repair antigen epitopes, can reduce the background, and can promote the specific binding between antibodies. Meanwhile, in the incubation process of the antibody, the membrane can be prevented from being broken or protein is prevented from being denatured due to drying; 3. The ready-to-use repairing liquid is nontoxic and harmless, is not easy to volatilize, has simple storage conditions and excellent tabletting effect by matching with a full-automatic immunohistochemical staining instrument, and can meet the requirements of clinical departments of hospitals on mass and high-quality pathological tabletting. Drawings The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate the invention and together with the embodiments of the invention, serve to explain the invention. In the drawings: FIG. 1 is a graph showing the effect of s