Detection of apoptosis in vivo using antibodies against caspase-induced neo-epitopes.
Cell death induction by apoptosis is an important process in the maintenance of tissue homeostasis as well as tissue destruction during various pathological processes. Consequently, detection of apoptotic cells in situ represents an important technique to assess the extent and impact of cell death in the respective tissue. While scoring of apoptosis by histological assessment of apoptotic cells is still a widely used method, it is likely biased by sensitivity problems and observed-based variations. The availability of caspase-mediated neo-epitope-specific antibodies offers new tools for the detection of apoptosis in situ. Here, we discuss the use of immunohistochemical detection of cleaved caspase 3 and lamin A for the assessment of apoptotic cells in paraffin-embedded liver tissue. Furthermore, we evaluate the effect of tissue pretreatment and antigen retrieval on the sensitivity of apoptosis detection, background staining and maintenance of tissue morphology.
Jakob
Sabine
S
Division of Immunopathology, Institute of Pathology, University of Bern, Murtenstrasse 31, 3010 Bern, Switzerland.
Corazza
Nadia
N
Diamantis
Eva
E
Kappeler
Andreas
A
Brunner
Thomas
T
D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000032:analysis* / Q000276:immunology; D000939:Epitopes; D005260:Female; D007150:Immunohistochemistry; D007797:Laminin / Q000032:analysis* / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D019014:fas Receptor / Q000502:physiology
D000939:Epitopes; D007797:Laminin; D019014:fas Receptor; C084098:laminin A; D053148:Caspase 3