Intraoperative quick immunohistochemical frozen-section staining method

Abstract

An intraoperative quick immunohistochemical frozen-section staining method comprises the following steps: placing an intraoperative fresh specimen in a freezing microtome for freezing and then slicing, wherein the section thickness is 5 microns, pasting a section on a glass slide, and carrying out air drying for 25-35 seconds; dropwise adding a primary antibody for incubation at 40 DEG C for 4 minutes; executing washing with PBS three times for 4-7 seconds each time; dropwise adding a secondary antibody for incubation at 40 DEG C for 4 minutes; executing washing with PBS three times for 4-7 seconds each time; dropwise adding a DAB reagent, and executing development for 1-2 minutes; executing staining with hematoxylin stain for 3-10 seconds. The staining process is not fixed, endogenous blocking is avoided, the incubation temperature is increased, the reaction time is shortened, the steps are omitted, the staining time is shortened, the total experiment time is greatly shortened, the whole process time needs about 12 minutes, the possibility of assisting the rapid frozen pathology diagnosis time is provided, the frozen diagnosis accuracy is improved, and the method has good repeatability and comparability with immunohistochemical results of paraffin.

Description

technical field [0001] The invention relates to the technical field of medical pathological detection, in particular to an immunohistochemical staining method for intraoperative frozen sections. Background technique [0002] Intraoperative frozen rapid pathology is to quickly give a preliminary pathological diagnosis (<30min) to a small part of the patient’s resected tissue under the microscope during the operation, and the preliminary pathological diagnosis will guide the doctor’s next surgical treatment plan, which has important clinical value. However, frozen diagnosis has certain limitations. Many tumors can’t be diagnosed only by intraoperative frozen staining technique, and the diagnosis can only be confirmed after postoperative paraffin specimen immunohistochemical examination. In this case, intraoperative rapid frozen pathological diagnosis is the Lost the clinical value of its specified surgical treatment plan. If reliable immunohistochemical staining results ca...

AI Technical Summary

Problems solved by technology

In the process of immunohistochemical staining, it is generally necessary to preserve the original structure of tissues and cells as much as possible to fix cells, to avoid degradation, autolysis and deformation of tissues and cells, and to lose various enzymes in cells and tissues. Activity, etc.; but fixed cells occupy a certain period of time during the staining process and some fixatives will cause cell shrinkage, damage to antigens, and damage to proteins, etc.