Simultaneous detection of TDT-mediated dUTP-biotin nick end-labeling (TUNEL)-positive cells and multiple immunohistochemical markers in single tissue sections

Biotechniques. 1995 Nov;19(5):800-5.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Apoptosis*
  • Biotin*
  • Cells, Cultured
  • DNA Nucleotidylexotransferase / metabolism*
  • Deoxyuracil Nucleotides / metabolism*
  • Female
  • Immunohistochemistry*
  • Mice
  • Neurons / physiology
  • Pregnancy
  • Silver Staining
  • Staining and Labeling / methods*
  • Telencephalon / cytology*
  • Telencephalon / embryology

Substances

  • Deoxyuracil Nucleotides
  • deoxyuridine triphosphate
  • Biotin
  • DNA Nucleotidylexotransferase