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