Simple and Rapid Method for Detection of Apoptotic Cells by Flow Cytometry
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概要
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We have developed a simple and rapid method for detection of apoptotic cells by flow cytometry -a modified nick end-labeling method. DNA fragments <I>in situ</I> 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 <I>in situ</I> 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.
- 日本組織細胞化学会の論文
- 1995-06-01
著者
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Hata Jun-ichi
Department Of Chemical Science And Engineering Graduate School Of Engineering Science Osaka Universi
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Yamada Taketo
Department Of Microbiology And Immunology School Of Medicine Keio University
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Yamada Taketo
Department Of Pathology Keio University Shool Of Medicine
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Hata Jun-ichi
Department Of Pathology Keio University Shool Of Medicine
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YAMADA Taketo
Department of Anesthesiology, School of Medicine, Keio University
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