Raman spectroscopy for DNA quantification in cell nucleus

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...

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Bibliographic Details
Published in:Cytometry. Part A 2015-01, Vol.87 (1), p.68-73
Main Authors: Okotrub, K. A., Surovtsev, N. V., Semeshin, V. F., Omelyanchuk, L. V.
Format: Article
Language:English
Subjects:
Animals
Cell Nucleus - ultrastructure
Chickens
DNA
DNA - analysis
Erythroblasts - ultrastructure
Feulgen cytometry
Humans
Lymphocytes - ultrastructure
nonstoichiometry
nucleus
Phosphates - chemistry
Raman spectroscopy
Rosaniline Dyes
Salamandridae
Spectrum Analysis, Raman
Zebrafish
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Summary: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
ISSN:1552-4922
1552-4930
DOI:10.1002/cyto.a.22585