Sahai Erik, Wyckoff Jeffrey, Philippar Ulrike, Segall Jeffrey E, Gertler Frank, Condeelis John
Tumour Cell Biology Laboratory, Cancer Research UK London Research Institute, London WC2A 3PX, UK.
BMC Biotechnol. 2005 May 23;5:14. doi: 10.1186/1472-6750-5-14.
The development of multiphoton laser scanning microscopy has greatly facilitated the imaging of living tissues. However, the use of genetically encoded fluorescent proteins to distinguish different cell types in living animals has not been described at single cell resolution using multiphoton microscopy.
Here we describe a method for the simultaneous imaging, by multiphoton microscopy, of Green Fluorescent Protein, Cyan Fluorescent Protein and collagen in vivo in living tumors. This novel method enables: 1) the simultaneous visualization of overall cell shape and sub-cellular structures such as the plasma membrane or proteins of interest in cells inside living animals, 2) direct comparison of the behavior of single cells from different cell lines in the same microenvironment in vivo.
Using this multi-fluor, multiphoton technique, we demonstrate that motility and metastatic differences between carcinoma cells of differing metastatic potential can be imaged in the same animal simultaneously at sub-cellular resolution.
多光子激光扫描显微镜的发展极大地促进了对活组织的成像。然而，尚未有研究利用多光子显微镜在单细胞分辨率下描述使用基因编码荧光蛋白来区分活体动物中不同细胞类型的情况。
在此，我们描述了一种通过多光子显微镜在活体肿瘤体内同时成像绿色荧光蛋白、青色荧光蛋白和胶原蛋白的方法。这种新方法能够：1）在活体动物体内同时可视化整体细胞形态和亚细胞结构，如质膜或细胞内感兴趣的蛋白质；2）直接比较来自不同细胞系的单个细胞在体内同一微环境中的行为。
使用这种多荧光、多光子技术，我们证明了具有不同转移潜能的癌细胞之间的运动性和转移差异能够在同一动物体内以亚细胞分辨率同时成像。
