Hideki Hatta | PubMed Health

PubMed2006

Freshly prepared immune complexes with intermittent microwave irradiation result in rapid and high-quality immunostaining.

We have previously reported that intermittent microwave irradiation shortened the primary or secondary antibody incubation time to 10 min in a special moist chamber. To achieve precise immunostaining within 1h, we attempted to generate a novel procedure, "freshly prepared immune complex with intermittent microwave irradiation (f-IC-M)". The advantage of this immunostaining procedure lies in a one-step incubation instead of primary and then secondary antibody application. In this study, we employed five primary antibodies to examine the efficiency and quality of this procedure. As expected, every primary antibody examined brought about precise immunostaining within 45 min for formalin-fixed, paraffin-embedded sections, and within 15 min for frozen sections. In addition, this procedure is able to generate double-immunoenzymatic staining with different enzyme-labeled primary antibodies if desired. As any combination of primary and secondary antibodies is possible by this one-step application, f-IC-M increases the efficiency of immunostaining without losing quality. Therefore, this procedure is able to rapidly provide diagnostic information to the pathologists.

Adenocarcinoma↗

PubMed2004

Improved 1-h rapid immunostaining method using intermittent microwave irradiation: practicability based on 5 years application in Toyama Medical and Pharmaceutical University Hospital.

Immunostaining depending on antigen-antibody specificity is the commonest approach for determining the localization of specific antigens in tissue sections. This procedure is applicable not only with frozen or specially fixed samples, but also has proved reliable with formalin-fixed paraffin-embedded tissue sections through improvement of antigen-retrieval. Immunostaining is thus firmly established as a tool for diagnostic pathology and in our institute multiple antibodies are applied for 13-15% of the cases examined, as well as H and E staining. With the standard approach, approximately 3 h is necessary from the beginning of deparaffinization till covering sections with the Envision system. We utilized intermittent microwave irradiation for 10 min during hybridization with primary and secondary antibodies in a special moist-chamber, to achieve all immunostaining steps within 1 h in 178 primary antibodies frequently used for diagnostic pathology. According to our 5 years experience, such microwave irradiation not only obtained significant specific staining for enhancing the specificity of antigen-antibody reactions, but also inhibited nonspecific binding. We present herein the details of the methodology and recommendations for its application with particular primary antibodies. This method can contribute to savings in time and energy, allowing pathologists to rapidly obtain diagnostic information.

Antibodies, Monoclonal↗