Simple and Rapid Method for Detection of Apoptotic Cells by Flow Cytometry.

• Yamada Taketo

  Department of Pathology, Keio University School of Medicine

• Hata Jun-ichi

  Department of Pathology, Keio University School of Medicine

Abstract

We have developed a simple and rapid method for detection of apoptotic cells by flow cytometry -a modified nick end-labeling method. DNA fragments in situ associated with apoptosis are detected by the nick-end labeling method (TUNEL method) in our new method. This method is well suited for easy identification of cell types and for in situ measurement of minute quantities of apoptotic cells. We present a modified method for nick-end labeling by flow cytometry (NELF) to detect apoptosis of both cultured cells and blood cells and also to analyze DNA fragments quantitatively and at the single-cell level. We modified the NELF method to simplify procedures and to save time. The protocol consists of 1) fixation with cold acetone, 2) transfer of biotiny lated nucleotide to the 3′-OH end with terminal deoxynucleotidyl transferase, and 3) incubation with streptoavidin-conjugated phycoerythrin to biotinylated nucleotide. This method may be useful in two-color analysis with a wide variety of antibodies.

Journal

• ACTA HISTOCHEMICA ET CYTOCHEMICA

  ACTA HISTOCHEMICA ET CYTOCHEMICA 28 (3), 277-280, 1995

  JAPAN SOCIETY OF HISTOCHEMISTRY AND CYTOCHEMISTRY

Keywords

• Apoptosis
• Nick end-labeling method
• DNA fragmentation
• Flow cytometry

Details

• CRID: 1390001204861842432
• NII Article ID: 10008603936
• NII Book ID: AA00508022
• DOI: 10.1267/ahc.28.277
• COI: 1:CAS:528:DyaK28XisFCl
• ISSN: 13475800
• ISSN: 00445991
• Web Site: http://www.jstage.jst.go.jp/article/ahc1968/28/3/28_3_277/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/1996061660
• Text Lang: en
• Data Source:
  • JaLC
  • Crossref
  • CiNii Articles
• Abstract License Flag: Disallowed