Ruthazer Edward S, Schohl Anne, Schwartz Neil, Tavakoli Aydin, Tremblay Marc, Cline Hollis T
Cold Spring Harb Protoc. 2013 Sep 1;2013(9):804-9. doi: 10.1101/pdb.top077156.
In vivo fluorescence imaging of cells in the developing nervous system is greatly facilitated in specimens in which cells are brightly but sparsely labeled. In this article, we describe a number of techniques that can be used for delivering fluorophore to neurons in the albino Xenopus laevis tadpole. Fluorescent dye or DNA that encodes a fluorescent protein can be delivered to single cells by electroporation. Alternatively, multiple cells can be labeled with fluorescent dye introduced by local iontophoresis or with plasmid DNA introduced by bulk electroporation. Technical considerations and analysis methods for time-lapse imaging in living tissue are also discussed.
在发育中的神经系统中，对于细胞被明亮但稀疏标记的标本，细胞的体内荧光成像得到了极大的便利。在本文中，我们描述了一些可用于将荧光团递送至白化非洲爪蟾蝌蚪神经元的技术。荧光染料或编码荧光蛋白的DNA可通过电穿孔递送至单个细胞。或者，多个细胞可用通过局部离子电渗法引入的荧光染料或通过批量电穿孔引入的质粒DNA进行标记。还讨论了活组织中延时成像的技术考量和分析方法。
