Optimization of estrogen receptor analysis by immunocytochemistry in random periareolar fine-needle aspiration samples of breast tissue processed as thin-layer preparations.

摘要: Immunostaining of estrogen receptor alpha (ER) in samples benign breast tissue obtained by random periareolar fine-needle aspiration (RPFNA) is a practical tool for cancer chemoprevention trials. The authors report an optimized method ER immunostaining use with thin-layer preparations modified Cytolyt-fixed acquired RPFNA. Samples and MCF-7 controls processed as were tested the effects antibody titer, antigen retrieval temperature (90 degrees or 115 C), buffer (20% nuclear decloaker, pH 9.3; 10 mM citrate buffer, 6), blocking solution (0.01% glucose oxidase 0.3% H2O2) on immunostaining. prevalence positively stained epithelial cells, mean intensity staining, composite score evaluated effect protocol. RPFNA cells using decloaker low-temperature had more ER-positive (P<0.0001) increased staining weighted indices (P<0.05) compared prepared high-temperature retrieval. Glucose comparison to hydrogen peroxide (P<0.04) when combined decloaker. Staining was negative all non-immune regardless combination retrieval, diluted commercial solution, step yielded optimal harvested