Raman spectroscopy of primary bovine aortic endothelial cells: a comparison of single cell and cell cluster analysis
Raman spectroscopy of primary bovine aortic endothelial cells: a comparison of single cell and cell cluster analysis
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DOI：
10.1007/s10856-011-4371-y
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发表时间：
2011-08-01
影响因子：
3.7
通讯作者：
Meenan, B. J.
中科院分区：
文献类型：
--
作者：
Boyd, A. R.;McManus, L. L.;Meenan, B. J.
有许多技术可以监测细胞及其环境之间的体外相互作用，包括分子、生物化学和免疫化学技术。用于分析细胞的传统技术通常需要固定或从基底裂解;然而，在需要将扩增的细胞培养物用于临床植入的情况下，使用这种破坏性方法是不可行的。先前的几项研究强调了拉曼光谱提供细胞内关键生化标记物的有用信息的潜力。因此，我们强调的能力，拉曼光谱与不同的激光光斑尺寸作为一种非侵入性的，快速的，和具体的方法来进行原位分析的原代牛主动脉内皮细胞（BAECs）。从在熔融石英基底上培养的单个活细胞和放置在熔融石英基底上的活细胞簇收集拉曼光谱，在532和785 nm处测量。所得结果显示DNA/RNA区域中的显著光谱差异，表明细胞质和细胞核的相对贡献。细胞簇的拉曼光谱显示苯丙氨酸峰（1004 cm（-1））强度的轻微变化，表明蛋白质贡献的变化。这些光谱还分别突出了来自其他细胞组分（例如蛋白质、脂质、核酸和碳水化合物）的贡献。
There are many techniques that allow in vitro interactions among cells and their environment to be monitored, including molecular, biochemical and immunochemical techniques. Traditional techniques for the analysis of cells often require fixation or lysis from substrates; however, use of such destructive methods is not feasible where the expanded cell cultures are required to be used for clinical implantation. Several studies have previously highlighted the potential of Raman spectroscopy to provide useful information on key biochemical markers within cells. As such, we highlight the capability of Raman spectroscopy with different laser spot sizes for use as a non-invasive, rapid, and specific method to perform in situ analysis of primary bovine aortic endothelial cells (BAECs). Raman spectra were collected from both individual live cells cultured on fused silica substrates and on clusters of live cells placed on fused silica substrates, measured at 532 and 785 nm. The results obtained show notable spectral differences in DNA/RNA region indicative of the relative cytoplasm and nucleus contributions. Raman spectra of cell clusters show slight variations in the intensity of the phenylalanine peak (1004 cm(-1)) indicating variations in protein contribution. These spectra also highlight contributions from other cellular components such as, proteins, lipids, nucleic acids and carbohydrates, respectively.