Ultrastructural and confocal laser scanning microscopic examination of tunel-postivie cells

Kenji Kimura, Hironobu Sasano, Tooru Shimosegawa, Katsuaki Kato, Tetsuya Noguchi, Shizue Mochizuki, Takashi Sawai, Masaru Koizumi, Takayoshi Toyota, Hiroshi Nagura

Research output: Contribution to journalArticlepeer-review

33 Citations (Scopus)

Abstract

TdT-mediated dUTP-biotin nick end labelling (TUNEL) has been widely used for detecting cells with DNA fragmentation or apoptotic cells. However, since the concept of apoptosis is based on cellular ultrastructure, it is important to identify the morphological features of TUNEL-positive cells. In this study, we performed TUNEL and electron microscopic observation on serial semithin and ultrathin sections of pancreas from bilaterally adrenalectomized rats with caerulein-induced pancreatitis. TUNEL-positive cells were identified with two different ultrastructural patterns. One was characteristic of apoptosis, with condensed nuclei, intact mitochondria, and zymogen granules. The other pattern was one of marked cellular degeneration, possibly representing the end stage of cell death. Cells which did not demonstrate these ultrastructuraI patterns were not labelled by the TUNEL method. The three-dimensional structure of TUNEL-positive cells was also investigated by confocal laser scanning microscopy (CLSM), which showed the apoptotic nuclei exhibited various three-dimensional structures. These results confirm the utility of the TUNEL method in detecting apoptosis; application of the technique reported in this study will contribute to the further characterization of individual TUNEL-positive cells.

Original languageEnglish
Pages (from-to)235-242
Number of pages8
JournalJournal of Pathology
Volume181
Issue number2
DOIs
Publication statusPublished - 1997

Keywords

  • Apoptosis
  • CLSM
  • Electron microscopy
  • Pancreas
  • Serial sections
  • TUNEL

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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