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Vital DNA staining and cell sorting by flow microfluorometry.

M J Lydon, K D Keeler, D B Thomas

    Journal of Cellular Physiology
    |February 1, 1980
    PubMed
    Summary
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    This study presents a method for sorting viable cells by DNA content using UV-activated fluorochromes like DAPI and Hoechst stains. The procedure is non-mutagenic and does not affect cell cloning efficiency, enabling new cell biology applications.

    Area of Science:

    • Cell Biology
    • Molecular Biology
    • Biotechnology

    Background:

    • Accurate cell sorting based on DNA content is crucial for various biological studies.
    • Existing methods may have limitations in terms of cell viability or specificity.

    Purpose of the Study:

    • To investigate a novel procedure for sorting viable cells based on their DNA content.
    • To evaluate the efficacy and safety of specific UV-activated fluorochromes for cell sorting.

    Main Methods:

    • Cells were stained with UV-activated fluorochromes: 4'6-diamidino-2-phenylindole (DAPI), Hoechst 33258, or Hoechst 33342.
    • Cell sorting was performed using a Fluorescence Activated Cell Sorter (FACS).
    • Vital staining properties and effects on cloning efficiency were assessed.

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    Main Results:

    • Hoechst 33342 demonstrated suitability as a vital stain across various cell types.
    • DAPI and Hoechst 33258 were identified as quantitative vital stains specifically for Chinese Hamster Ovary (CHO) cells.
    • The cell sorting procedure did not negatively impact cloning efficiency.
    • The vital staining concentrations used were found to be non-mutagenic.

    Conclusions:

    • The developed procedure enables effective sorting of viable cells by DNA content using specific fluorochromes.
    • This technique offers a non-mutagenic and reliable method for cell analysis.
    • Potential applications in cell biology research are significant, allowing for advanced cell population studies.