[An intergral method for quantitative cell DNA content measurements by digital microphotography].
Quantitative nuclei DNA content measurement based on Feulgen reaction and the analysis of CCD images was tested. The measurements were performed in monochorome CCD option (650 X 514 pixels) with the wavelength 551 nm. The linear dependence of photomatrix elements signals on the falling light was shown with the use of multigraded light absorption filter. The optimal microscope and camera setting and an approach for elimination of the optic blur were found. We have shown that the proper Feulgen fluorescence does not affect our measurements. Densitometric DNA content measurements of blood cells of four vertebrate species (Gallus domesticus, Danio rerio, Homo sapiens and Rana arvalis) showed good consistence to the literature data (http://www.genomesize.com). The precision of our approach is comparable to the other known methods. Current improvement of CCD technical parameters and wide usage of CCD cameras in biological applications gives perspectives for the suggested approach.
Omel'ianchuk
L V
LV
Semeshin
V F
VF
Alekseeva
A L
AL
Pal'chikova
I G
IG
Zhimulev
I F
IF
D000818:Animals; D001773:Blood Cells / Q000737:chemistry; D002467:Cell Nucleus / Q000737:chemistry; D003592:Cytophotometry / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000032:analysis*; D003720:Densitometry; D006801:Humans; D010781:Photography; D012394:Rosaniline Dyes / Q000737:chemistry; D012680:Sensitivity and Specificity
C007722:Feulgen stain; D012394:Rosaniline Dyes; D004247:DNA