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NASA/American Cancer Society High-Resolution Flow Cytometry Project - II. Effect of pH and DAPI concentration on dual parametric analysis of DNA/DAPI fluorescence and electronic nuclear volume.
- Source :
-
Cytometry [Cytometry] 2001 Jan 01; Vol. 43 (1), pp. 12-5. - Publication Year :
- 2001
-
Abstract
- Background: In the present paper, we describe the effect of 4', 6-diamidino-2-phenylindole (DAPI) dihydrochloride concentration and pH on the resolution of DNA distribution histograms generated by dual-parametric simultaneous analysis of DNA content and electronic nuclear volume (ENV).<br />Methods: Nuclei from tissue culture cell lines and frozen human solid tumors were isolated in nuclear isolation media containing different concentrations of DAPI, at various pH levels, and analyzed on a NASA/American Cancer Society (ACS) flow cytometer. Samples stained with propidium iodide/hypotonic citrate and analyzed in a Coulter XL flow cytometer were used for comparison.<br />Results: Nuclei stained with DAPI concentration of 1-3 microg/ml, pH 6.0, gave the best resolution for the detection of the near-diploid and near-tetraploid populations. Simultaneous use of ENV and DAPI/DNA fluorescence under these conditions identified subpopulations that otherwise could not be detected by DNA analysis alone.<br />Conclusions: Staining at 1-3 microg/ml DAPI, pH 6.0, was optimal for the detection of aneuploid populations, especially the near-diploid and/or near-tetraploid populations in human tumors.
- Subjects :
- Aneuploidy
Animals
Cell Line
DNA metabolism
Erythrocytes cytology
Female
Fluorescence
Humans
Hydrogen-Ion Concentration
Male
Mice
Neoplasms genetics
Neoplasms pathology
Ploidies
Propidium metabolism
United States
American Cancer Society
Cell Nucleus pathology
DNA analysis
Flow Cytometry methods
Indoles metabolism
United States National Aeronautics and Space Administration
Subjects
Details
- Language :
- English
- ISSN :
- 0196-4763
- Volume :
- 43
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Cytometry
- Publication Type :
- Academic Journal
- Accession number :
- 11122479