Comparison of vibrational spectroscopy to biochemical and flow cytometry methods for analysis of the basic biochemical composition of mammalian cells
Comparison of vibrational spectroscopy to biochemical and flow cytometry methods for analysis of the basic biochemical composition of mammalian cells
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DOI：
10.1117/1.2400213
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发表时间：
2006-11-01
影响因子：
3.5
通讯作者：
Freyer, James P.
中科院分区：
文献类型：
--
作者：
Mourant, Judith R.;Dominguez, Jorge;Freyer, James P.
我们对从完整细胞的红外和拉曼光谱中获得的细胞生化成分与使用标准提取和化学分析（包括 NMR）以及固定细胞的流式细胞术测定进行的测量进行了广泛的比较。对大鼠成纤维细胞癌变模型进行测量，该模型由正常细胞和致瘤细胞组成，测定为指数生长和平台期培养物。蛋白质、DNA、RNA、脂质和糖原含量的估计值是从之前的出版物中获得的，其中振动光谱适合代表蛋白质、DNA、RNA、脂质和糖原的一组基础光谱。绝对细胞组成的拉曼光谱估计与独立的生化和流式细胞术测定非常相似。红外光谱对蛋白质、脂质和糖原给出了类似的结果，但低估了 DNA 含量，同时高估了 RNA 水平。当检查指数培养物和平台期培养物中的生化浓度比率时，在所有情况下，拉曼光谱结果与独立方法相同（在误差范围内）。独立方法证实了先前使用振动光谱法报道的由于致瘤和增殖状态导致的相对生化成分的一些变化。这些结果表明振动光谱可以提供哺乳动物细胞生化成分的可靠估计。 (c) 2006 年光电仪器工程师协会。
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. (c) 2006 Society of Photo-Optical Instrumentation Engineers.