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Journal of Histochemistry and Cytochemistry, Vol. 49, 623-630, May 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

A New Rapid Immunohistochemical Staining Technique Using the EnVision Antibody Complex

Ulrike Kämmerera, Michaela Kappa, Andrea Maria Gasselb, Thomas Richterc, Christian Tankd, Johannes Dietla, and Peter Rucke
a Department of Obstetrics and Gynecology, University of Wuerzburg, Wuerzburg, Germany
b Department of Pathology, University of Wuerzburg, Wuerzburg, Germany
c Department of Pathology, Technical University of Munich, Munich, Germany
d DAKO Corporation, Hamburg, Hamburg, Germany
e Department of Pathology, University of Tuebingen, Tuebingen, Germany

Correspondence to: Ulrike Kämmerer, Dept. of Obstetrics and Gynecology, University of Würzburg, Josef-Schneider Str. 4, D-97080 Würzburg, Germany. E-mail: frak057@mail.uni-wuerzburg.de

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)

Key Words: immunohistochemistry, frozen section diagnosis, EnVision


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