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同步辐射傅里叶变换红外(FTIR)分析单个活细胞在细胞周期中的进展。

Synchrotron Fourier transform infrared (FTIR) analysis of single living cells progressing through the cell cycle.

机构信息

Centre for Biospectroscopy, School of Chemistry, Monash University, 3800, Victoria, Australia.

出版信息

Analyst. 2013 Jul 21;138(14):3891-9. doi: 10.1039/c3an00316g. Epub 2013 Jun 12.

Abstract

The application of FTIR spectroscopy to disease diagnosis requires a thorough knowledge of the spectroscopy associated with the cell cycle to discern disease markers from normal cellular events. We have applied synchrotron FTIR spectroscopy to monitor cells at different phases of the cell cycle namely G1, S and G2 phases. By applying Principal component analysis (PCA) from three independent trials we show clustering on a 2-dimensional scores plots (PC1 versus PC2) from cell spectra only two hours apart within the cell cycle. The corresponding PCA Loadings Plots indicate the clustering is primarily based on changes to the overall concentration of nucleic acids, proteins and lipids. During the first ten hours post mitosis, cells are observed to increase in protein and decrease in both lipid and nucleic acid concentration. During the synthesis phase, (beginning 9-11 hours post-mitosis) the PCA Loadings Plots show the accumulation of lipids within the cell as well the duplication of the genome as evidenced by strong DNA contributions. In the 4-6 hours following the synthesis phase, the cells once again accumulate protein while the relative nucleic acid and lipid concentrations decrease. These results, in comparison to previous studies on dehydrated cells, show previously unresolvable biochemical information as well as highlighting the advantages of FTIR spectroscopy applied to single living cells.

摘要

傅里叶变换红外(FTIR)光谱学在疾病诊断中的应用需要深入了解与细胞周期相关的光谱学,以从正常细胞事件中辨别疾病标志物。我们已经将同步辐射 FTIR 光谱学应用于监测处于不同细胞周期阶段的细胞,即 G1、S 和 G2 期。通过对三个独立实验的主成分分析(PCA),我们在细胞光谱的二维得分图(PC1 对 PC2)上显示出聚类,这些细胞光谱仅在细胞周期内相隔两小时。相应的 PCA 载荷图表明聚类主要基于核酸、蛋白质和脂质的总体浓度变化。在有丝分裂后最初的十个小时内,观察到细胞内蛋白质增加,同时脂质和核酸浓度降低。在合成期(有丝分裂后 9-11 小时开始),PCA 载荷图显示细胞内脂质的积累以及基因组的复制,这可以通过 DNA 的强烈贡献来证明。在合成期结束后的 4-6 小时内,细胞再次积累蛋白质,而相对的核酸和脂质浓度降低。与以前对脱水细胞的研究相比,这些结果显示了以前无法解决的生化信息,同时也突出了 FTIR 光谱学应用于单个活细胞的优势。

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