1887

Abstract

The DNA and protein content of individual cells were measured at a rate of 10 cells per second with a sensitive microscope-based flow cytometer. DNA and protein were quantified by measuring the fluorescence from cells stained with a combination of the DNA-binding drugs Mithramycin and ethidium bromide and by scattered light, respectively. Separate experiments demonstrated that the light scatter signal was proportional to protein content. Dual parameter histograms (fluorescence/scattered light) of bacterial cultures gave detailed pictures of changes dependent upon the growth conditions and of the cell cycle kinetics. Effects of antibiotics could be readily detected and characterized after a few hours. The results demonstrate that flow cytometry is a promising method for application in experimental and clinical microbiology.

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/content/journal/micro/10.1099/00221287-129-4-973
1983-04-01
2021-10-19
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References

  1. Bailey J. E., Fazel-Madjlessi J., Mcquitty U. IN., Lee L. Y., Allred J. C., Oro J. A. 1977; Characterization of bacterial growth by means of flow microfluorometry. Science 198:1175–1176
    [Google Scholar]
  2. Barlogie B., Spitzer G., Hart J. S., Johnston D. A., BÜchner T., Schmann J., Drewinko B. 1976; DNA histograms of human hemopoietic cells. Blood 48:245–258
    [Google Scholar]
  3. Boye E., Krisch R. E. 1980; Induction and repair of DNA double-strand breaks in bacteriophage lambda superinfecting E. coli. International Journal of Radiation Biology 37:119–133
    [Google Scholar]
  4. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  5. Lindmo T. 1981; Protein synthesis as a function of protein content in exponentially growing NHIK 3025 cells studied by flow cytometry and cell sorting. Experimental Cell Research 133:237–245
    [Google Scholar]
  6. Lindmo I., Steen H. B. 1979; Characteristics ol a simple, high-resolution flow cytometer based on a new flow configuration. Biophysical Journal 28:33–44
    [Google Scholar]
  7. Paau A. S., Lee D., Cowles J. R. 1977a; Comparison of nucleic acid content in free-living and symbiotic Rhizobium meliloti by flow microfluorometry. Journal of Bacteriology 129:1156–1158
    [Google Scholar]
  8. Paau A. S., Cowles J. R., Oro J. A. 1977b; Flow microfluorometric analyses of Escherichia coli, Rhizobium meliloti, and Rhizobium japonicum in different stages in the growth cycle. Canadian Journal of Microbiology 23:1165–1169
    [Google Scholar]
  9. Steen H. B. 1980; Further developments of a microscope-based flow cytometer: light scatter detection and excitation intensity compensation. Cytometry 1:26–31
    [Google Scholar]
  10. Steen H. B., Boye E. 1980; Bacterial growth studied by flow cytometry. Cytometry 1:32–36
    [Google Scholar]
  11. Steen H. B., Boye E. 1981; Escherichia coli growth studied by dual-parameter flow cytophotometry. Journal of Bacteriology 145:1091–1094
    [Google Scholar]
  12. Steen H. B., Lindmo T. 1979; Flow cytometry: a high resolution instrument for everyone. Science 204:403–404
    [Google Scholar]
  13. Ward D. C., Reich E., Goldberg I. H. 1965; Base specificity in the interaction of polynucleotides with antibiotic drugs. Science 149:1259–1263
    [Google Scholar]
  14. Wechsler J. A., Gross J. D. 1971; Escherichia coli mutants temperature-sensitive for DNA synthesis. Molecular and General Genetics 113:273–284
    [Google Scholar]
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