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DOI: 10.1369/jhc.4B6563.2005
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Journal of Histochemistry and Cytochemistry
Volume 53 (8): 1033-1036, 2005
Copyright ©The Histochemical Society, Inc.

BRIEF REPORT

DAPI Fluorescence in Nuclei Isolated from Tumors

Awtar Krishan and Payal D. Dandekar

Division of Experimental Therapeutics, Radiation Oncology Department, Sylvester Cancer Center, University of Miami, School of Medicine, Miami, Florida

Correspondence to: Dr. Awtar Krishan, Division of Experimental Therapeutics (R-71), University of Miami, School of Medicine, PO Box 01690, Miami, FL 33136. E-mail: akrishan{at}med.miami.edu

In DNA histograms of some human solid tumors stained with nuclear isolation medium–4,6-diamidino-2-phenylindole dihydrochloride (NIM-DAPI), the coefficient of variation (CV) of the G0/G1 peak was broad, and in nuclear volume vs DNA scattergrams, a prominent slope was seen. To determine the cause for this, nuclei from frozen breast tumors were stained with NIM-DAPI and analyzed after dilution or resuspension in PBS. In two-color (blue vs red) analysis, most of the slope and broad CV was due to red fluorescence of nuclei stained with NIM-DAPI, which was reduced on dilution or resuspension in PBS, resulting in elimination of the slope and tightening of the CV.

(J Histochem Cytochem 53:1033–1036, 2005)

Key Words: tumors • nuclear volume • flow cytometry • DAPI • DNA content


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