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FLOW CYTOMETRIC DISCRIMINATION OF MITOTIC NUCLEI BY RIGHT-ANGLE LIGHT SCATTER (JOURNAL VERSION)
Citation:
Zucker, R., K. Elstein, R. Easterling, AND E. Massaro. FLOW CYTOMETRIC DISCRIMINATION OF MITOTIC NUCLEI BY RIGHT-ANGLE LIGHT SCATTER (JOURNAL VERSION). U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-88/094.
Description:
Flow cytometry has been used to demonstrate alterations in protein, RNA, and DNA content of cells as they traverse the cell cycle. Employing fluorescein isothiocynate (FITC) to stain protein and propidium iodide (PI) to stain nucleic acids, multiple regions within the G1 and G2 phases of the cell cycle, in addition to the M phase, can be distinguished. Cytograms of the 90 degree light scatter signal vs. P1 fluorescence were remarkably similar to those of FITC fluorescence vs. P1 fluorescence, suggesting a relationship between 90 degree light scatter and protein content. M-phase cells can be distinguished from G2-phase cells on cytograms of 90 degree light scatter vs. P1 fluorescence. However, the percent of mitotic cells obtained by this technique is approximately 30% less than that found by light microscopic analysis. Flow cytometric parameters of nuclei prepared by nonionic detergent (NP40) lysis in Dulbecco's PBS, Vindelov's buffer, or Pollack's hypotonic EDTA/Tris buffer were compared. The best resolution of mitotic cells were obtained in Pollack's buffer. However, the stainability of the M-phase cells is reduced and they are located in the G2/M region of the single-parameter histogram. (Copyright (c) 1988 Alan R. Liss, Inc.)