Brandenburg Cheryl, Poulopoulos Alexandros
Department of Pharmacology and Physiology, University of Maryland School of Medicine, Baltimore, MD, USA.
Methods Mol Biol. 2025;2910:263-276. doi: 10.1007/978-1-0716-4446-1_16.
This protocol outlines plasmid delivery via in utero electroporation, rapid tissue clearing using CUBIC, and light sheet microscopy with optimizations for endogenous fluorescent protein imaging to label and image neuronal cells and their projections in their native topographic context of the intact developing rodent cerebellum. This technique enables the study of neuronal migration, circuit architecture, and connectivity of cerebellar neurons, particularly focusing on Purkinje cells.
本方案概述了通过子宫内电穿孔进行质粒递送、使用CUBIC进行快速组织透明化以及光片显微镜检查，并针对内源性荧光蛋白成像进行了优化，以在完整发育的啮齿动物小脑的天然地形背景下标记和成像神经元细胞及其投射。该技术能够研究小脑神经元的迁移、电路结构和连接性，尤其侧重于浦肯野细胞。
