Department of Genetics and Development, Columbia University Irving Medical Center, New York, NY 10032, USA.
Department of Translational Medical Science, University of Campania Luigi Vanvitelli, 80131 Napoli, Italy.
STAR Protoc. 2022 Jun 10;3(2):101427. doi: 10.1016/j.xpro.2022.101427. eCollection 2022 Jun 17.
Despite being among the largest neurons in the mammalian brain, Purkinje cells are difficult to visualize and trace via immunofluorescence because their dendritic arbors extend through several cerebellar layers. This protocol describes a two-antibody strategy we developed to study Purkinje cell morphology in mice. With it, one can reconstruct three-dimensional images of Purkinje cells at single-neuron resolution across multiple layers. The substantially improved image quality reveals subtle defects, enabling more meaningful morphological analysis. For complete details on the use and execution of this protocol, please refer to Gennarino et al. (2015).
尽管浦肯野细胞是哺乳动物大脑中最大的神经元之一，但由于其树突分支延伸穿过几个小脑层，因此通过免疫荧光很难对其进行可视化和追踪。本方案描述了我们开发的一种用于研究小鼠浦肯野细胞形态的双抗体策略。使用该策略，可以在多个层以单神经元分辨率重建浦肯野细胞的三维图像。显著提高的图像质量揭示了细微的缺陷，从而实现更有意义的形态分析。有关该方案使用和实施的完整详细信息，请参见 Gennarino 等人（2015 年）。
