Simultaneous in situ detection of mRNA and apoptotic cells by combined hybridization and TUNEL.
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.
Sträter
J
J
Institute of Pathology, University of Ulm, Germany.
Walczak
H
H
Krammer
P H
PH
Möller
P
P
D017209:Apoptosis; D006801:Humans; D017403:In Situ Hybridization; D012333:RNA, Messenger / Q000235:genetics*; D019014:fas Receptor / Q000235:genetics
D012333:RNA, Messenger; D019014:fas Receptor