Kinetics of epitope retrieval techniques for unmasking cytokeratins in bovine prostatic tissues after different formaldehyde fixation times.

PORTIANSKY EL; MASSONE AR; GIMENO EJ

APPLIED IMMUNOHISTOCHEMISTRY

LIPPINCOTT WILLIAMS & WILKINS

Lugar: Philadelphia; Año: 1997 vol. 5 p. 194 - 201

1062-3345

lmmunohistochemistry is a powerful too! for tissue diagnosisand research. Usually, routine formali n-fixed, paraffinembeddedtissue blocks are the only materials available. However,exposure to formaldehyde produces progressive loss oftissue epitopes. In the past few years, physical methods forantigen unmasking have been developed. In this study, we analyzedthe immunoreactivity of bovine prostatic basal cell cytokeratins13 and 16 after fixation in 4% formaldehyde for 6, 12,and 24 hours, and 7, 15, and 30 days. An evident fall-off inimmunoreactivity as a function of fixation time was observed.Microwave-citrate (MWC), pressure cooker (PC), and ultrasoundretrieval (USR) techniques enhanced immunostain ingcompared with routine immunohistochemistry. Statistically, theUSR method was shown to be superior in terms of immunostainedarea and optical densily (OD) at all evaluated fixationtimes. Additionally, in the USR procedure, retrieval exposuretime was very short and the heating effect was negligible; thus,loss of sections and structural damage to the tissue did notoccur.