Quantitative Analysis On The Influence Of Tissue Antigen In The Key Procedure Of Immunohistochemistry

Objectives: To investigate the effects of the pre-treatment steps in the immunohistochemistry on the protein antigen and find out the factors that influence the antigenicity,and to quantitatively analyze the influence of the reagent used for tissue treatment on the antigenicity.Methods: With diagnosed breast cancer and estrogen receptor immunohistochemical staining for positive tissues were produced for frozen sections and paraffin sections as the samples for immunohistochemical staining respectively,with phosphate buffer substituting the first antibody as negative control,finally analyze the staining results;The serum samples with positive hepatitis B surface antigen was simulated to the tissue processing procedures of tissue samples.The 10% neutral formalin,absolute ethanol and xylene,paraffin wax(paraffin simulating the melting point temperature),xylene and absolute ethanol were added to serum;normal saline was added as control group.Then we use the antibody absorption test when the reaction has been fully completed;the concentration of hepatitis B surface antibody was determined after the reaction,and the data were statistically analyzed by statistical software.Results: There was no difference in the results of immunohistochemical staining of frozen sections and paraffin sections with antigen.The results of immunohistochemical staining of paraffin sections without antigen retrieval were negative and there is difference between the former two;There is no difference in antigenicity between the 10% neutral formalin treated serum and control group;but there were differences between the serum that were treated with absolute alcohol and control group(P<0.5),without antigen retrieval the loss of antigenicity was >90%.Conclusions:The results showed that the absolute alcohol reagent had the most significant effect on the antigenicity in the IHC pretreatment procedures.Therefore,the IHC technique perhaps is only suitable for the detection of large quantity and strong antigenicity antigen in tissues.