SIMPLE METHOD FOR PRETREATMENT OF TISSUE-SECTIONS FOR THE DETECTION OF APOPTOSIS BY IN-SITU END-LABELING AND IN-SITU NICK TRANSLATION

Aims-To overcome the problems associated with proteolytic pretreatment of tissue sections for the detection of apoptosis. Methods-Formalin f ixed, paraffin wax embedded tissue sections of reactive lymph nodes an d biopsy specimens of Burkitt lymphoma were pretreated by pressure coo king for the detection of apoptosis using the in situ end-labelling an d in situ nick translation methods. Results-The results achieved with the in situ end-labelling and nick translations methods were compared with those obtained using a novel anti-apoptosis specific protein (ASP ) antibody. The staining patterns generated using the three methods we re similar and consistent, although the ASP antibody seemed to be more sensitive and detected higher numbers of apoptotic cells within secti ons. Conclusions-Pressure cooking is advocated as an alternative metho d to proteolytic enzyme digestion for pretreating paraffin wax section s. It is reliable, inexpensive, reduces the need to optimise pretreatm ent variables for different tissues, and permits double immunostaining of sections.