A New Rapid Immunohistochemical Staining Technique Using the EnVision Antibody Complex

TLDR

Rapid immunohistochemistry could aid intraoperative frozen‑section diagnosis. The study examined whether the EnVision system could be adapted for rapid frozen‑section immunostaining. Forty‑five primary antibodies were applied to frozen sections that were acetone‑fixed, air‑dried, and incubated for 3 min each with primary antibody, EnVision complex, and chromogen at 37 °C. The modified EnVision method produced specific staining with 38 of 45 antibodies within 13 min, demonstrating broad applicability for frozen‑section diagnosis. J Histochem Cytochem 49:623–630 (2001).

Abstract

Rapid immunohistochemical investigation, in addition to staining with hematoxylin and eosin, would be useful during intraoperative frozen section diagnosis in some cases. This study was undertaken to investigate whether the recently described EnVision system, a highly sensitive two-step immunohistochemical technique, could be modified for rapid immunostaining of frozen sections. Forty-five primary antibodies were tested on frozen sections from various different tissues. After fixation in acetone for 1 min and air-drying, the sections were incubated for 3 min each with the primary antibody, the EnVision complex (a large number of secondary antibodies and horseradish peroxidase coupled to a dextran backbone), and the chromogen (3,3′diaminobenzidine or 3-amino-9-ethylcarbazole). All reactions were carried out at 37C. Specific staining was seen with 38 antibodies (including HMB-45 and antibodies against keratin, vimentin, leukocyte common antigen, smooth muscle actin, synaptophysin, CD34, CD3, CD20, and prostate-specific antigen). A modification of the EnVision method allows the detection of a broad spectrum of antigens in frozen sections in less than 13 min. This method could be a useful new tool in frozen section diagnosis and research. (J Histochem Cytochem 49:623–630, 2001)