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Simultaneous in situ detection of mRNA and apoptotic cells by combined hybridization and TUNEL

J Strater, H Walczak, PH Krammer and P Moller

Institute of Pathology, University of Ulm, Germany.

We established a new method to allow simultaneous in situ detection of mRNA expression and apoptotic DNA fragmentation in paraffin-embedded tissue sections. We used human thymic tissue to perform in situ hybridization with a digoxigenin-labeled CD95 (APO-1/Fas) ligand (CD95L)-specific probe followed by TdT-mediated biotin dUTP nick end- labeling (TUNEL) of apoptotic DNA fragments. Bound probes were visualized by an immunogold-silver enhancement technique and fragmented DNA was detected with a streptavidin-peroxidase system. This double labeling technique produced a distinct, dark cytoplasmic staining of CD95L mRNA-expressing cells and an intense red nuclear precipitate in apoptotic cells or bodies. This technique will be a useful tool for microtopographical analysis of apoptosis-related gene expression.

Volume 44, Issue 12, pp. 1497-1499, 12/01/1996
Copyright © 1996 by The Histochemical Society


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