Nanotechnology and Integrated Bioengineering Centre, University of Ulster, Shore Road, Newtownabbey, Antrim, BT37 0QB, Northern Ireland.
J Mater Sci Mater Med. 2011 Aug;22(8):1923-30. doi: 10.1007/s10856-011-4371-y. Epub 2011 Jun 14.
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.
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、生化和免疫化学技术。传统的细胞分析技术通常需要从基质中固定或溶解细胞；然而，在需要将扩展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在提供细胞内关键生化标记物有用信息方面的潜力。因此，我们强调了不同激光光斑大小的拉曼光谱在作为一种非侵入性、快速和特定方法原位分析原代牛主动脉内皮细胞 (BAEC) 方面的能力。从培养在熔融石英基底上的单个活细胞和放置在熔融石英基底上的活细胞簇中收集拉曼光谱，在 532 和 785nm 处进行测量。所得结果显示 DNA/RNA 区域的光谱差异明显，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（1004cm-1）强度略有变化，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成分（如蛋白质、脂质、核酸和碳水化合物）的贡献。
