DAPI fluorescence in nuclei isolated from tumors.
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.
Krishan
Awtar
A
Division of Experimental Therapeutics, Radiation Oncology Department, Sylvester Cancer Center, University of Miami, School of Medicine, Miami, FL 33136, USA. akrishan@med.miami.edu
Dandekar
Payal D
PD
D000818:Animals; D001943:Breast Neoplasms / Q000737:chemistry* / Q000648:ultrastructure; D002467:Cell Nucleus / Q000737:chemistry* / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D007211:Indoles
D004273:DNA, Neoplasm; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI