EFFECT OF HEAT-INDUCED ANTIGEN RETRIEVAL PROCEDURE ON DNA FLUORESCENT STAINING QUALITY IN HISTOLOGIC SECTIONS

Abstract

The heat-induced epitope retrieval procedure is an important step in the procedure of immunofluorescent staining of tissues fixed in formalin-containing solutions. Generally, apart from the detection of the examined antigen, the staining of cell nuclei w ith appropriate fluorescent dyes is also carried out during the morphological studies. This procedure is used to identify the cells position and form, the phase of the mitotic cycle, the size of the nucleus and the nucleolus, the chromatin condition. The aim of our study is to investigate the effects of heat-induced epitope retrieval (HIER) procedure on the quality of DNA staining by fluorescent dyes DAPI and 7-Aminoactinomycin D. We have found that the potential effect of the HIER procedure on the stability and intensity of cell nuclei fluorescence staining should be taken into account in selecting a protocol for immunofluorescence antigen identification during a morphological study. Data obtained in the course of this study should be taken into account when carrying out a multicolor fluorescence and confocal laser microscopy.

Keywords

nuclear staining dyes / fluorochromes / fluorescent dyes / heat-induced epitope retrieval

RIGHTS & PERMISSIONS

Kolos E.A., Korzhevskii D.E.