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 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.
['Okotrub|K A|KA|', 'Surovtsev|N V|NV|', 'Semeshin|V F|VF|', 'Omelyanchuk|L V|LV|']
D000818:Animals D002467:Cell Nucleus D002645:Chickens D004247:DNA D004900:Erythroblasts D006801:Humans D008214:Lymphocytes D010710:Phosphates D012394:Rosaniline Dyes D012455:Salamandridae D013059:Spectrum Analysis, Raman D015027:Zebrafish
Raman spectroscopy DNA nucleus Feulgen cytometry nonstoichiometry