Ultrastructural Analysis of a Mouse Brain Section Using Transmission Electron Microscopy

Abstract

Source: Lutz, D., et.al. Assessment of Ultrastructural Neuroplasticity Parameters After In Utero Transduction of the Developing Mouse Brain and Spinal Cord. J. Vis. Exp. (2019).

This video demonstrates the preparation and transmission electron microscopy (TEM) analysis of a brain section from a mouse pup. The protocol starts with osmium tetroxide-fixed tissue embedded in resin, followed by precise trimming to isolate the region of interest and ultramicrotomy to produce semithin and ultrathin sections, which are visualized using light microscopy and TEM, respectively.

Protocol

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

1. Selection of Ultrastructural Neuroplasticity Parameters for Quantitative Analysis

1. Mapping the area of interest
   1. Choose an area of interest (e.g., hippocampus or cerebellum) and localize the area in the section atlas by choosing....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Light microscope Basic DM E	Leica	-	4x (N.A. 0.1 ∞/-), 10x (N.A. 0.22 ∞/0.17), 40x (N.A. 0.65 ∞/0.17), 100x (N.A. 1.25 ∞/0.17) objectives
Mosquito hemostatic forceps (12.5cm, curved)	FST	13010-12
Nickel grids, 200 mesh	Ted Pella	1GC200
Razor blades	Schick	87-10489
Technovit 4004 two components glue	Kulzer
Thin vibrating razor blade device	Krup	-	with Szabo thin blades
Toluidine blue	Sigma-Aldrich	89640
Transmission electron microscope C20	Phillips	-	up to 200 kV
Ultracut E	Reichert-Jung	-	ultramicrotome
Araldite	CIBA-GEIGY	23857.9	resin for embedding of tissue
Osmium (VIII)-oxid	Degussa	73219