Generation of Tilted Amygdala Slices from a Mouse Brain

Abstract

Source: Bosch, D., et al. Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices. J. Vis. Exp. (2016)

The video demonstrates the generation of tilted amygdala slices from a mouse brain. The brain is positioned at an angle on the vibratome specimen stage to obtain slices containing the amygdala and axons from the medial prefrontal cortex that form synaptic connections with the amygdala. The brain slices are maintained at the physiological temperature to preserve the tissue for further experiments.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.

1. Preparation of Acute Slices

1. Prepare artificial cerebrospinal fluid (ACSF), cutting solution, tools (scissors, scalpel, forceps, spatula, Pasteur pipette), and agar blocks.
   Note: 1 L of artificial cerebrospinal fluid (ACSF) is required per experiment, and is prepared by dissolving chemicals in double-distilled H₂O as previously published. The cutting solution is prepared by supplementing 200 ml of ACSF with 0.87 ml of 2 M MgSO₄ stock solution.

Results

Figure 1. Stereotactic Injections, Preparation of Acute Brain Slices, and Visualization of Presynaptic Fibers. (A, B) Stereotactic virus injection. A) Picture of anesthetized mouse placed in a stereotactic frame with skull exposed and t.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
artificial cerebrospinal fluid (ACSF)
internal patch solutions
Magnesium Sulfate Heptahydrate	Roth, Germany	P027.1	prepare 2M stock solution in purified water
Slicer, Microm HM650V	Fisher Scientific, Germany	920120
Cooling unit for tissue slicer, CU65	Fisher Scientific, Germany	770180
Sapphire blade	Delaware Diamond Knives		custom order, inquire with company
Stereoscope, SZX2-RFA16	Olympus, Japan
Patch microscope, BX51WI	Olympus, Japan
Bath temperature controler, TC05	Luigs & Neumann, Germany	200-100 500 0145
Cellulose nitrate filterpaper for interface chamber	Satorius Stedim Biotech, Germany	13006--50 ACN