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Raman spectroscopy for DNA quantification in cell nucleus
Author(s) -
Okotrub K. A.,
Surovtsev N. V.,
Semeshin V. F.,
Omelyanchuk L. V.
Publication year - 2015
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22585
Subject(s) - dna , raman spectroscopy , densitometry , feulgen stain , cytometry , nuclear dna , raman scattering , chemistry , biophysics , microbiology and biotechnology , cell , biology , biochemistry , optics , physics , gene , mitochondrial dna
Here we demonstrate the feasibility of a novel approach to quantify DNA in cell nuclei. This approach is based on spectroscopy analysis of Raman light scattering, and avoids the problem of nonstoichiometric binding of dyes to DNA, as it directly measures the signal from DNA. Quantitative analysis of nuclear DNA contribution to Raman spectrum could be reliably performed using intensity of a phosphate mode at 1096 cm −1 . When compared to the known DNA standards from cells of different animals, our results matched those values at error of 10%. We therefore suggest that this approach will be useful to expand the list of DNA standards, to properly adjust the duration of hydrolysis in Feulgen staining, to assay the applicability of fuchsines for DNA quantification, as well as to measure DNA content in cells with complex hydrolysis patterns, when Feulgen densitometry is inappropriate. © 2014 International Society for Advancement of Cytometry