Combined Labelled and Label-free SERS Probes for Triplex Three-dimensional Cellular Imaging.
Combined Labelled and Label-free SERS Probes for Triplex Three-dimensional Cellular Imaging.
复制标题
用于三重三维细胞成像的组合标记和无标记 SERS 探针
DOI：
10.1038/srep19173
复制
发表时间：
2016-01-19
影响因子：
4.6
通讯作者：
Hu J
中科院分区：
文献类型：
--
作者：
Chen Y;Bai X;Su L;Du Z;Shen A;Materny A;Hu J
细胞是复杂的化学系统，其中不同细胞位置的分子组成和特定的细胞内化学相互作用决定了生物学功能。我们的研究目标是对复杂的化学过程（如细胞凋亡）进行原位无损表征。在这里，我们提出的结果，同时和三维成像的双细胞器（细胞核和膜）在单个HeLa细胞通过标记或无标记的表面增强拉曼光谱（SERS）。当使用荧光显微镜时，这种有标记和无标记的成像组合是不可能的。SERS技术用于立体描述细胞核的内在化学性质，以及在高度共聚焦条件下膜上叶酸（FA）和促黄体生成素释放激素（LHRH）的精确定位。我们还报道了细胞核以及膜受体蛋白在凋亡过程中的时间依赖性变化，通过统计多变量方法进行分析。多重三维SERS成像技术可以同时对活细胞进行时间（真实的时间）和空间（三维空间中的多个细胞器和分子）成像，从而为生物医学亚细胞水平上的二维/三维示踪提供了一种新的、有吸引力的方法。
Cells are complex chemical systems, where the molecular composition at different cellular locations and specific intracellular chemical interactions determine the biological function. An in-situ nondestructive characterization of the complicated chemical processes (like e.g. apoptosis) is the goal of our study. Here, we present the results of simultaneous and three-dimensional imaging of double organelles (nucleus and membrane) in single HeLa cells by means of either labelled or label-free surface-enhanced Raman spectroscopy (SERS). This combination of imaging with and without labels is not possible when using fluorescence microscopy. The SERS technique is used for a stereoscopic description of the intrinsic chemical nature of nuclei and the precise localization of folate (FA) and luteinizing hormone-releasing hormone (LHRH) on the membrane under highly confocal conditions. We also report on the time-dependent changes of cell nuclei as well as membrane receptor proteins during apoptosis analyzed by statistical multivariate methods. The multiplex three-dimensional SERS imaging technique allows for both temporal (real time) and spatial (multiple organelles and molecules in three-dimensional space) live-cell imaging and therefore provides a new and attractive 2D/3D tracing method in biomedicine on subcellular level.