Visualization of a Mouse Brain Choroid Plexus Using Scanning Electron Microscopy

Abstract

Source: Van Wonterghem, E., et al. Microdissection and Whole Mount Scanning Electron Microscopy Visualization of Mouse Choroid Plexus. J. Vis. Exp. (2022)

This video demonstrates the preparation and imaging of choroid plexus tissue from a mouse brain using scanning electron microscopy (SEM). The process includes tissue fixation, dehydration, and coating with a conductive platinum layer to stabilize and prepare the sample for imaging. SEM captures detailed images of epithelial cells with microvilli, providing insights into the tissue’s structural features.

Protocol

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

1. Morphological analysis of choroid plexus (CP) tissue using scanning electron microscopy (SEM)

CAUTION: Toxic solutions are used in the following processing steps. It is recommended to perform the sample preparation in a fume hood.

1. Perform sample preparation for SEM as described below.
   1. Transfer the fresh isolated CP into freshly made fixation solution containing 2% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M Na-cacodylate buffer (pH 7.4). Incubate overnight at 4 &....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Aluminium specimen mounts	EM Sciences	75220
Cacodylate buffer	EM Sciences	11652
Carbon adhesive tabs -12 mm	EM Sciences	77825-12
Critical point dryer	Bal-Tec	CPD030
Crossbeam 540	Zeiss		SEM system
Forceps	Fine Science Tools GmbH	91197-00
Glutaraldehyde	EM Sciences	16220
Osmium Tetroxide	EM Sciences	19170
Paraformaldehyde	Sigma-Aldrich	P6148
Phosphate buffered saline (PBS)	Lonza	BE17-516F
Platinum	Quorum	Q150T ES	PBS without Ca⁺⁺, Mg⁺⁺ or phenol red; sterile filtered
Specimen Basket Stainless Steel	EM Sciences	70190-01