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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.

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Sparse Labeling, Rapid Clearing, and Native Fluorescence Light Sheet Imaging in the Developing Rodent Cerebellum

  • 实验方案

  • 发表日期：

    2025年

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  • Cheryl Brandenburg,
  • Alexandros Poulopoulos

摘要

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.