Visualizing the Synaptogenesis of Cerebellar Granule Neurons at Different Developmental Stages

Abstract

Source: Chan, U., et al., Utilizing In Vivo Postnatal Electroporation to Study Cerebellar Granule Neuron Morphology and Synapse Development. J. Vis. Exp. (2021).

This video demonstrates the analysis of cerebellar granule neuron development by imaging pre-fixed brain tissue slices from mice labeled with a fluorescent protein, capturing high-resolution z-stack images with a confocal microscope, and measuring structural parameters including neurite length, dendritic claw formation, surface area, and volume to study synaptogenesis.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Morphological analyses of cerebellar granule neurons or CGNs - three-dimensional (3D) reconstruction and surface area and cellular volume

1. Image single electroporated CGNs on a confocal microscope at 63x objective with a 2x zoom, taking z-stack images at 0.5 µm per stack. Image one cell per image window to allow for easy image analysis and re....

Results

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Imaris	Bitplane
Zeiss 780 Upright Confocal	Zeiss	780