Department of Biomedical Engineering, National Taiwan University, Taipei, Taiwan.
Department of Ophthalmology, Chang Gung Memorial Hospital, Linkou, Taiwan.
Methods Mol Biol. 2020;2150:131-140. doi: 10.1007/7651_2019_227.
Multiphoton microscopy allows long-term direct visualization of cells in live animals due to its low photodamage. When coupled with fluorescence protein targeting and second harmonic generation signals from natural collagen as contrast, multiphoton microscopy enables intravital tracing of cells while providing structural information from the extracellular matrix. Compared with conventional histological analysis, it can bring new insight into the cell dynamics in stem cell research. Here, we demonstrate cell imaging and tracing at a single cell resolution in the cornea, skin, and hair follicles using multiphoton microscopy in transgenic mice of which specific cell populations are tagged with fluorescent proteins.
多光子显微镜由于其低光损伤特性，可实现活体内细胞的长期直接可视化。当与荧光蛋白靶向和天然胶原的二次谐波产生信号结合使用作为对比时，多光子显微镜可实现细胞的活体示踪，同时提供细胞外基质的结构信息。与传统的组织学分析相比，它可以为干细胞研究中的细胞动力学提供新的见解。在这里，我们展示了在具有荧光蛋白标记的特定细胞群体的转基因小鼠中，使用多光子显微镜在角膜、皮肤和毛囊中以单细胞分辨率进行细胞成像和示踪。
