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Related Experiment Video

Updated: May 14, 2025

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy

Published on: October 8, 2013

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

Cheryl Brandenburg¹, Alexandros Poulopoulos²

¹Department of Pharmacology and Physiology, University of Maryland School of Medicine, Baltimore, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)

|April 12, 2025

Summary

This study presents a method for visualizing neuronal development in the rodent cerebellum using in utero electroporation, tissue clearing, and advanced microscopy. This technique allows detailed observation of neuronal migration and circuit formation, especially for Purkinje cells.

Keywords:

CUBIC clearing Cerebellum Light sheet imaging Plasmid electroporation Projection mapping Purkinje cell labeling

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Last Updated: May 14, 2025

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Area of Science:

Neuroscience
Developmental Biology
Microscopy

Background:

Studying the developing cerebellum requires methods to visualize neuronal structure and connectivity in vivo.
Existing techniques may have limitations in resolution, depth, or preserving native tissue context.

Purpose of the Study:

To develop and optimize a comprehensive protocol for imaging neuronal development in the rodent cerebellum.
To enable detailed analysis of neuronal migration, circuit architecture, and connectivity.

Main Methods:

Plasmid delivery via in utero electroporation.
Rapid tissue clearing using CUBIC (Clear, Unobtanium, Bond, Immersion, Chemical).
Light sheet microscopy with optimized endogenous fluorescent protein imaging.

Main Results:

Successful labeling and imaging of neuronal cells and their projections in the intact developing rodent cerebellum.
Visualization of neuronal migration patterns and the topographic context of cerebellar circuits.
Detailed imaging of Purkinje cells and their axonal/dendritic arbors.

Conclusions:

This integrated technique provides a powerful tool for studying cerebellar development and circuitry.
It facilitates research into neuronal migration, circuit formation, and connectivity with high fidelity.
The protocol is particularly valuable for investigating Purkinje cell development and function.