SIMULTANEOUS DETECTION OF TDT-MEDIATED DUTP-BIOTIN NICK END-LABELING (TUNEL)-POSITIVE CELLS AND MULTIPLE IMMUNOHISTOCHEMICAL MARKERS IN SINGLE TISSUE-SECTIONS

We have modified the terminal deoxynucleotidyl transferase (TDT)-media ted dUTP-biotin nick end-labeling (TUNEL) method to permit the immunog old-silver intensification detection of apoptotic cells in tissue sect ions. Such sections can subsequently be processed for multi-labeling f luorescence microscopy, thus permitting the simultaneous detection of silver-positive apoptotic cells and three additional florescent signal s. In this report, we combined TUNEL labeling with the fluorescent det ection of bromodeoxyuridine-positive proliferating cells, growth-assoc iated protein-43 (GAP-43) immunoreactive neurons and bisbenzimide-posi tive pyknotic cells in the embryonic mouse nervous system. This combin ation of stains allowed the simultaneous detection of proliferative, d ifferentiated and apoptotic cells in the developing nervous system.