Flow cytometric discrimination of mitotic nuclei by right-angle light scatter

✍ Scribed by Robert M. Zucker; Kenneth H. Elstein; Robert E. Easterling; Edward J. Massaro

• Publisher: John Wiley and Sons
• Year: 1988
• Tongue: English
• Weight: 547 KB
• Volume: 9
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/cyto.990090307

✦ Synopsis

Flow cytometry has been used to demonstrate alterations in protein, RNA, and DNA content of cells as they traverse the cell cycle. Employing fluorescein isothiocyanate (FITC) to stain protein and propidium iodide (PI) to stain nucleic acids, multiple regions within the GI and Gz phases of the cell cycle, in addition to the M phase, can be distinguished. In this study, cytograms of the 90" light scatter signal vs. PI fluorescence were remarkably similar to those of FITC fluorescence vs. PI fluorescence, suggesting a relationship between 90" light scatter and protein content. M-phase nuclei can be distinguished from Gzphase nuclei on cytograms of 90" light scatter vs. PI fluorescence. However, the percentage of mitotic nuclei obtained by this technique is less than that found by light microscopic analysis. Flow cytometric parameters of nuclei prepared by nonionic detergent W " ~O ) lysis in Dulbecco's PBS, Vindelov's buffer, or Pollack's hypotonic EDTAfTris buffer were compared. The best resolution of mitotic nuclei was obtained in Pollack's buffer. However, the stainability of the M-phase nuclei is reduced, and the nuclei are located in the late S/Gz region of the single-parameter histogram.