TUNEL, Hoechst and immunohistochemistry triple-labelling: an improved method for detection of apoptosis in tissue sections--an update.
We describe an updated protocol which allows the combination of the previously described TUNEL and Hoechst 33342 double-labelling (G. Whiteside, N. Cougnon, S.P. Hunt, R. Munglani, An improved method for the detection of apoptosis in tissue sections and cell culture, using the TUNEL technique combined with Hoechst stain, Brain Res. Protocols, 2, 1998, 160-164 [3]), with immunohistochemistry. The inclusion of the immunohistochemistry step allows identification of apoptosis in distinct cell types by using cell specific markers. It also permits investigation into the proteins expressed in cells undergoing apoptotic cell death. We have evaluated the triple staining in sections of post-axotomy neonatal dorsal root ganglia.
Whiteside
G
G
Department of Anaesthesia, University of Cambridge, Level 4 Addenbrookes Hospital, Hills Road, Cambridge CB2 2QQ, UK. gtw20@medschl.cam.ac.uk
Munglani
R
R
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001562:Benzimidazoles; D005456:Fluorescent Dyes; D005727:Ganglia, Spinal / Q000737:chemistry* / Q000473:pathology; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D008867:Microtomy; D051381:Rats
D001562:Benzimidazoles; D005456:Fluorescent Dyes; C017807:bisbenzimide ethoxide trihydrochloride