An evaluation of DNA fluorochromes, staining techniques, and analysis for flow cytometry. I. Unperturbed cell populations

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Show simple item record Taylor, IW Milthorpe, BK 2010-05-28T09:45:25Z 1980
dc.identifier.citation Journal of Histochemistry and Cytochemistry, 1980, 28 (11), pp. 1224 - 1232
dc.identifier.issn 0022-1554
dc.identifier.other C1UNSUBMIT en_US
dc.description.abstract Several preparative techniques (detergent treatment, ethanol fixation, and hypotonic cell lysis), DNA fluorochromes, and methods of numerical analysis (planimetric or curve-fitting) were compared for the estimation of cell-cycle kinetic parameters (G1, S, G2 + M) by flow cytometry. In addition, coefficients of variation (CV), relative fluorescence, and G1/chicken erythrocyte (CRBC) ratios were measured and the effects of the proportion of cycling cells and cellular RNA content were examined. DNA-fluorochromes were ranked by relative fluorescence: 4,6-diamidino-2-phenylindole > ethidium bromide/mithramycin > Hoechst 33342 > mithramycin > ethidium bromide > acridine orange ≃ propidium iodide. The first four (DNA-specific stains) gave lower CVs than the remainder (DNA intercalators). Detergent treatment also increased relative fluorescence and slightly lowered CVs. Comparable results were obtained for the kinetic parameters independently of stain or staining procedure; intercalating dyes with cells of a high RNA content not treated with RNAse and acridine orange being exceptions. Of the two methods of numerical analysis, the planimetric technique was more consistent. Although highly consistant G1/CRBC ratios were obtained for any one stain, independently of staining procedures, variations between stains were noted. It is suggested that the detergent treatment in combination with DNA-specific stains provide optimal results.
dc.language eng
dc.title An evaluation of DNA fluorochromes, staining techniques, and analysis for flow cytometry. I. Unperturbed cell populations
dc.type Journal Article
dc.parent Journal of Histochemistry and Cytochemistry
dc.journal.volume 11
dc.journal.volume 28
dc.journal.number 11 en_US
dc.publocation United States en_US
dc.identifier.startpage 1224 en_US
dc.identifier.endpage 1232 en_US SCI.Medical and Molecular Biosciences en_US
dc.conference Verified OK en_US
dc.for 0601 Biochemistry and Cell Biology
dc.personcode 105631
dc.percentage 100 en_US Biochemistry and Cell Biology en_US
dc.classification.type FOR-08 en_US
dc.edition en_US
dc.custom en_US en_US
dc.location.activity en_US
dc.description.keywords Cell-cycle kinetics; Fluorescent dyes; Stains and staining; Cytology; Flow cytometry. en_US
dc.description.keywords Cell-cycle kinetics
dc.description.keywords Fluorescent dyes
dc.description.keywords Stains and staining
dc.description.keywords Cytology
dc.description.keywords Flow cytometry.
pubs.embargo.period Not known
pubs.organisational-group /University of Technology Sydney
pubs.organisational-group /University of Technology Sydney/Faculty of Science
pubs.organisational-group /University of Technology Sydney/Strength - Health Technologies

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