Is the central dogma of flow cytometry true: That fluorescence intensity is proportional to cellular dye content?

M. Kerker, M. A. Van Dilla, A. Brunsting, J. P. Kratohvil, P. Hsu, D. S. Wang, J. W. Gray, R. G. Langlois

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

Measurements and theoretical calculations of fluorescent emission from four samples of polystyrene microspheres (diameter 0.92, 1.63, 1.90 and 4.18 m̈m) containing the same fluorescent dye show a general dependence upon particle size, emission angle, and polarization conditions. However, for the excitation and detection conditions used in flow cytometry, the relative fluorescent intensities measured for the four particle sizes are proportional to the dye content to +10% accuracy, independent of particle size. Accordingly, the central dogma of flow cytometry ‘that fluorescence is proportional to cellular dye content’ is valid to this accuracy for these solid, highly refractive polymer particles. Most mammalian cells are much less refractive, therefore, should conform more closely to the central dogma.

Original languageEnglish (US)
Pages (from-to)71-78
Number of pages8
JournalCytometry
Volume3
Issue number2
DOIs
StatePublished - Sep 1982

Keywords

  • cellular dye content
  • Flow cytometry
  • fluorescence angular distribution
  • fluorescence emission theory
  • fluorescence polarization
  • fluorescent microspheres
  • fluorescent signal intensity
  • particle size

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

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  • Cite this

    Kerker, M., Van Dilla, M. A., Brunsting, A., Kratohvil, J. P., Hsu, P., Wang, D. S., Gray, J. W., & Langlois, R. G. (1982). Is the central dogma of flow cytometry true: That fluorescence intensity is proportional to cellular dye content? Cytometry, 3(2), 71-78. https://doi.org/10.1002/cyto.990030202