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Strategies for choosing a DNA stain for flow cytometry of metaphase chromosomes

Conference ·
OSTI ID:7142989

Requirements for flow cytometry of metaphase chromosomes stained with three DNA-specific fluorescent dyes (Hoechst 33258, Chromomycin A/sub 3/, and ethidium bromide) are reviewed. Fluorescence properties of these three stains when bound to mitotic cells or to chromosomes in suspension are measured and compared with fluorescence properties when bound to DNA in solution. Conditions are given for high resolution flow cytometry of Chinese hamster chromosomes stained with each of the fluorophors, and histograms are presented which exhibit differences in relative peak position and area. Energy transfer fluorescence between two DNA stains is presented as a potentially useful new parameter for flow cytometry of chromosomes and is illustrated by fluorescence energy transfer from Chromomycin A/sub 3/ to ethidium bromide when simultaneously bound to hamster mitotic cells.

Research Organization:
California Univ., Livermore (USA). Lawrence Livermore Lab.
DOE Contract Number:
W-7405-ENG-48
OSTI ID:
7142989
Report Number(s):
UCRL-79331; CONF-770417-1
Country of Publication:
United States
Language:
English

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