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Journal of Biomedical Optics

Comparison of vibrational spectroscopy to biochemical and flow cytometry methods for analysis of the basic biochemical composition of mammalian cells
Author(s): Judith R. Mourant; Jorge R. Dominguez; Susan R. Carpenter; Kurt R. Short; Tamara R. Powers; Ryszard Michalczyk; Nagapratima Kunapareddy; Anabel Guerra; James P. Freyer
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Paper Abstract

We have conducted an extensive comparison of cellular biochemical composition obtained from infrared and Raman spectra of intact cells with measurements using standard extraction and chemical analysis (including NMR), and flow cytometric assay on fixed cells. Measurements were conducted on a rat fibroblast carcinogenesis model consisting of normal and tumorigenic cells assayed as exponentially growing and plateau-phase cultures. Estimates of protein, DNA, RNA, lipids, and glycogen amounts were obtained from a previous publication in which vibrational spectra were fit to a set of basis spectra representing protein, DNA, RNA, lipids, and glycogen. The Raman spectral estimates of absolute cellular composition were quite similar to the independent biochemical and flow cytometric assays. The infrared spectra gave similar results for protein, lipid, and glycogen but underestimated the DNA content while overestimating the RNA level. When ratios of biochemical concentrations in exponential and plateau-phase cultures were examined, the Raman spectroscopic results were the same, within errors, as the independent methods, in all cases. Several changes in relative biochemical composition due to tumorigenic and proliferative status previously reported using vibrational spectroscopy were confirmed by the independent methods. These results demonstrate that vibrational spectroscopy can provide reliable estimates of the biochemical composition of mammalian cells.

Paper Details

Date Published: 1 November 2006
PDF: 11 pages
J. Biomed. Opt. 11(6) 064024 doi: 10.1117/1.2400213
Published in: Journal of Biomedical Optics Volume 11, Issue 6
Show Author Affiliations
Judith R. Mourant, Los Alamos National Lab. (United States)
Jorge R. Dominguez, Los Alamos National Lab. (United States)
Susan R. Carpenter, Los Alamos National Lab. (United States)
Kurt R. Short, Los Alamos National Lab. (United States)
Tamara R. Powers, Los Alamos National Lab. (United States)
Ryszard Michalczyk, Los Alamos National Lab. (United States)
Nagapratima Kunapareddy, Los Alamos National Lab. (United States)
Anabel Guerra, Los Alamos National Lab. (United States)
James P. Freyer, Los Alamos National Lab. (United States)

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