Institute of Photonics & Bio-medicine, School of Science, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China; Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1, Yamadaoka, Suita City, Osaka 565-0871, Japan.
Institute of Photonics & Bio-medicine, School of Science, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.
Spectrochim Acta A Mol Biomol Spectrosc. 2020 Jan 5;224:117438. doi: 10.1016/j.saa.2019.117438. Epub 2019 Jul 29.
Raman microspectroscopy as a non-invasive and label-free technique was applied to diagnose the early stage differentiation of mouse embryonic stem cells. The differentiated and undifferentiated embryonic bodies (EBs) were cultured using handing drop method by the control of Leukemia Inhibitory Factor (LIF). Raman spectra of the periphery cells of differentiated EBs (PrE cells) and those of the interior of undifferentiated EBs (ES cells) were obtained to diagnose the stem cells of different differentiation. It was found from the spectra that the protein content increased as the cells differentiated. Principal component analysis (PCA) was carried out to further analyze the differences between ES cells and PrE cells. The first three principle components contained 98.19% from the total variance. Characteristic bands of ES and PrE cells were chosen to acquire Raman images of two cells according to the results of PCA. In the Raman images, PrE cells had a clear and bright outline in the peripheral areas while ES cells were difficult to identify, this could be a distinct characteristic to discriminate them. The result of the Raman images was consistent with the biological agreement that the differentiated cells were distributed around the periphery.
拉曼微光谱分析作为一种非侵入式和无标记技术，被应用于诊断小鼠胚胎干细胞的早期分化。通过白血病抑制因子（LIF）的控制，使用手动滴注法培养分化和未分化的胚胎体（EBs）。获得分化 EB 的外周细胞（PrE 细胞）和未分化 EB 内部（ES 细胞）的拉曼光谱，以诊断不同分化的干细胞。从光谱中发现，随着细胞分化，蛋白质含量增加。进行主成分分析（PCA）以进一步分析 ES 细胞和 PrE 细胞之间的差异。前三个主成分包含总方差的 98.19%。根据 PCA 的结果，选择 ES 和 PrE 细胞的特征带，以获取两种细胞的拉曼图像。在拉曼图像中，PrE 细胞在外周区域有清晰明亮的轮廓，而 ES 细胞难以识别，这可能是区分它们的明显特征。拉曼图像的结果与生物学协议一致，即分化细胞分布在周边。
