Generating a Coronal Brain Slice Exposing the Amygdala Regions from a Mouse Brain

In This Article

Abstract

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

This video demonstrates the preparation of thin coronal brain slices from a murine brain, exposing the amygdala regions rich in synaptic connections from the medial prefrontal cortex. The slices are then recovered in oxygenated artificial cerebrospinal fluid to maintain neuronal viability.

Protocol

1. Preparation of Acute Slices

Prepare 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. As previously published, ACSF is prepared by dissolving chemicals in double-distilled H2O. The cutting solution is prepared by supplementing 200 ml of ACSF with 0.87 ml of 2 M MgSO₄ stock solution.
Oxygenate ACSF and cutting solution throughout the experiment with 95% O₂ and 5% CO₂.
Deeply anesthetize the mouse using a small animal anesthesia mach....

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Results

Stereotactic virus injection process in brain; diagram of injection sites and viral constructs layout.
Figure 1. Stereotactic Injections, Preparation of Acute Brain Slices, and Visualization of Presynaptic Fibers. (A, B) Stereotactic virus injection. A) Picture of an anesthetized mouse placed in a stereotactic frame with the skull expo.......