Label-Free Live-Cell Imaging with Confocal Raman Microscopy
Label-Free Live-Cell Imaging with Confocal Raman Microscopy
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DOI：
10.1016/j.bpj.2011.12.027
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发表时间：
2012-01-18
影响因子：
3.4
通讯作者：
Schlegel, Juergen
中科院分区：
文献类型：
--
作者：
Klein, Katharina;Gigler, Alexander M.;Schlegel, Juergen
共焦拉曼光谱是一种非侵入性的替代既定的细胞成像方法，因为它不需要化学固定，荧光标记物的使用，或基因工程。特别是，单活细胞，高分辨率成像共焦拉曼显微镜是可取的，因为它允许进一步的实验，有关个别调查的细胞。然而，为了从光谱数据中获得有意义的图像，必须识别数据集中的细胞成分。使用免疫荧光图像作为参考，我们得到拉曼光谱特征的信息措施，以确定细胞成分，如细胞核，内质网，高尔基体，和线粒体。提取的签名使我们能够生成相当于传统的（免疫）荧光图像与三个以上的细胞成分的时间表示，单独利用拉曼光谱信息。
Confocal Raman spectroscopy is a noninvasive alternative to established cell imaging methods because it does not require chemical fixation, the use of fluorescent markers, or genetic engineering. In particular, single live-cell, high-resolution imaging by confocal Raman microscopy is desirable because it allows further experiments concerning the individually investigated cells. However, to derive meaningful images from the spectroscopic data, one must identify cell components within the dataset. Using imnnunofluorescence images as a reference, we derive Raman spectral signatures by means of information measures to identify cell components such as the nucleus, the endoplasmic reticulum, the Golgi apparatus, and mitochondria. The extracted signatures allow us to generate representations equivalent to conventional (immuno)fluorescence images with more than three cell components at a time, exploiting the Raman spectral information alone.