The Effects On HE Staining,Immunohistochemistry And Molecular Detection Of Breast Tumor Tissue Processed By Intraoperative Frozen Procedure

Breast cancer has become one of the common cancers that threaten women’s health at this stage.Taking into account the overall condition of the patient,immunohistochemistry(IHC)is the main pathological diagnosis.Before IHC,there is a certain degree of tissue The treatment methods are generally conventional sectioning and frozen sectioning.Conventional sectioning is also called paraffin sectioning.After the tissue is fixed,it is embedded in paraffin,and then sectioned according to the needs,and the corresponding index detection is performed or after the dewaxing scraping.For genetic testing,the advantage is that it can be stored for a long time and can completely determine the condition of each cell in the tissue.Frozen sections are often combined with intraoperative rapid examinations.Generally,clinicians will determine the specificity of the tumor(malignant tumor range,pathological diagnosis or lymph node status etc.)during surgery.The advantage is that it is fast,but in the process of freezing the tissue,The production of ice crystals will cause damage to the cell structure.This experiment compares conventional sections,frozen sections,and “frozen leftover”tissues of the same tissue to observe whether different processing methods have an impact on breast cancer detection indicators,and compare the two in terms of DNA fragmentation,to study the degradation of DNA.1.Patient data collection and specimen processing.The clinical pathological data of breast cancer patients who received both conventional and frozen sections from the Department of Pathology of Hubei Cancer Hospital from August 2020 to October 2020 were collected,and pathological classification was performed in accordance with the2003 WHO breast tumor histological standards.The clinical staging was based on The8 th edition of the American Joint Committee on Cancer Clinical Analysis Standard in2018,the medical record data comes from the pathology and molecular pathology medical record system.Find the corresponding wax block according to the pathology number,sort it out,re-slice it,divide it into the experimental group and the control group according to different procedures,re-interpret and record.2.HE,IHC staining and FISH detection.HE staining was used to check the nucleus and cell membrane of the tissue.It was found that the overall staining of the experimental group was poor,the cell volume was significantly shrunk,and the nucleolus and nuclear membrane were not clear,which affected the interpretation of the slice.IHC staining detects the expression of ER,PR,HER-2,KI-67,E-ca,and P120 on the tissues.It is found that the indicators of ER,PR and Ki67 nuclear staining are accurate in the positive positioning of the experimental group and the control group,but the experimental group is stained The intensity is deeper than the control group(p<0.05),and some tissues also have cytoplasmic staining;the E-cadherin and p120 membranes are stained accurately,and the staining intensity is statistically significant compared with the control group(p<0.001).The staining intensity of E-cadherin in the group was stronger than that of the control group,but p120 was weaker than that of the control group.The HER-2 staining location was accurate.Although the experimental group was stained deeper than the control group,the difference was not statistically significant(p>0.05).According to the performance of the main detection indicators for breast cancer,determine the FISH test object.FISH is mainly HER-2/CEP17.Check the amplification of the HER-2 gene.It is found that the experimental group and the control group have differences in digestion time,ut there is no difference in the quality of the entire production.3.DNA extraction,purification and fragmentation detection.Frozen sections,experimental group and control group sections were subjected to scraping dewaxing purification treatment to detect the fragmentation degree of the three types of sections at100bp-300 bp,and to identify the effects of different processing methods on DNA degradation.It was found that the CT values of the 100 bp,200bp and 300 bp fragments of the experimental group and the control group were not statistically different,but compared with the frozen slice group,the latter had a smaller CT value for the 300 bp fragment,indicating that the DNA quality of the frozen slice group was better than that of the experimental group And the control group,the degradation degree is lower.Through the above research,it is found that the conventional sections and conventional sections of "frozen leftover tissue" are generally consistent with the immunohistochemical detection part,but there are still certain differences in the degree of staining,and most of the frozen leftover tissues have a membrane-positive phenomenon.It is speculated that it is related to the freshness of tissue preservation;from the perspective of DNA degradation,the difference is relatively large.Generally speaking,it does not affect the pathological diagnosis of cytology,and has no effect on tumor classification,indicating that the wax block has undergone a rapid intraoperative freezing process.It can be used for molecular pathology detection,but it is not suitable for IHC and HE staining to determine tissue type and index expression.