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)-mediated dUTP-biotin nick end-labeling (TUNEL) method to permit the immunogold-silver intensification detection of apoptotic cells in tissue sections. Such sections can subsequently be processed for multi-labeling fluorescence microscopy, thus permitting the simultaneous detection of silver-positive apoptotic cells and three additional fluorescent signals. In this report, we combined TUNEL labeling with the fluorescent detection of bromodeoxyuridine-positive proliferating cells, growth-associated protein-43 (GAP-43) immunoreactive neurons and bisbenzimide-positive pyknotic cells in the embryonic mouse nervous system. This combination of stains allowed the simultaneous detection of proliferative, differentiated and apoptotic cells in the developing nervous system.
Tornusciolo
D R
DR
Washington University School of Medicine, St. Louis, MO, USA.
Schmidt
R E
RE
Roth
K A
KA
D000818:Animals; D017209:Apoptosis; D001710:Biotin; D002478:Cells, Cultured; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism*; D003856:Deoxyuracil Nucleotides / Q000378:metabolism*; D005260:Female; D007150:Immunohistochemistry; D051379:Mice; D009474:Neurons / Q000502:physiology; D011247:Pregnancy; D016622:Silver Staining; D013194:Staining and Labeling / Q000379:methods*; D013687:Telencephalon / Q000166:cytology* / Q000196:embryology
D003856:Deoxyuracil Nucleotides; C027078:deoxyuridine triphosphate; D001710:Biotin; D004253:DNA Nucleotidylexotransferase