Integral method for measuring the quantity of cellular DNA content by digital microphotography

Autor: I. G. Palchikova, Leonid V. Omelyanchuk, A. L. Alekseeva, Igor F. Zhimulev, V. F. Semeshin
Rok vydání: 2010
Předmět:
Zdroj: Cell and Tissue Biology. 4:305-308
ISSN: 1990-5203
1990-519X
Popis: Quantitative measurements of nuclear DNA content based on Feulgen reaction and the analysis of CCD images has been proposed. The measurements were performed in the monochrome CCD option (650 × 514 pixels) with a wavelength of 551 nm. The linear dependence of photomatrix element signals on the falling light was shown with a multigrade light absorption filter. The optimal microscope and camera settings and an approach for elimination of the optic blur are proposed. It was found that the contribution of background fluorescence of Feulgen-stained nuclei into the measurements was negligible. Densitometric measurements of the DNA content in blood cells of four vertebrate species (Gallus domesticus, Danio rerio, Homo sapiens, Rana arvalis) were consistent with the literature data. The precision of our approach is comparable to other known cytometry methods (http://www.genomesize.com). The current improvement of CCD technical parameters and the widespread use of CCD cameras in biological applications give perspectives for the development of the suggested approach for measuring the quantity of cellular DNA.
Databáze: OpenAIRE
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