MICROSPECTROPHOTOMETRIC QUANTITATION OF IMMUNOHISTOCHEMICAL REACTIONS BY IMAGE ANALYSIS

Immunohistochemistry is a technique by immunological reactions to identify proteins in different biological samples. As a result leave a product in histological preparations, visible under a microscope when using a colour reaction. This is an indicator of the presence of the specific molecule and cellular metabolic activity represents a more or less specific. In the literature have used different methods to estimate the intensity of this reaction trying to get a reproducible value. With our microarray methodology (Cabrini RL et al.) we performed a spectral curve for the final product of the reaction of benzidine using as reaction control of a case inmunohistochemistry HER2 positive breast adenocarcinoma. Images were obtained from a single field of the preparation in a Zeiss MPM 800 with an objective x 40/0, 75 different light wavelengths. The maximum absorption was 0.55 optical density at 530nm per pixel. The measurement was performed in 10 cases of carcinomas STA1. The methodology presented in this study showed different values at predetermined fixed areas.