Obtaining Horizontal Mouse Hippocampal Brain Slices

Abstract

Source: Van Hoeymissen, E., et al. Horizontal Hippocampal Slices of the Mouse Brain. J. Vis. Exp. (2020)

This video demonstrates a detailed protocol for preparing horizontal hippocampal brain slices from mice, focusing on preserving the integrity of hippocampal fiber pathways. The process involves carefully bisecting and securing the brain, using a vibratome for slicing, and maintaining optimal tissue conditions with a carbogenated fluid, resulting in hippocampal brain slices ready for further experimentation.

Protocol

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

1. Preparation of high-sucrose slice solution and artificial cerebrospinal fluid (ACSF)

1. Prior to experimental day
   1. Prepare 1 L of 10x slice pre-solution with laboratory grade Type 1 water containing (in mM): 25 KCl, 20 CaCl₂, 10 MgSO₄, 12.5 KH₂PO₄ (Table 1). In order to prevent calcium phosphate precipitation, slowly mix the chemicals in a beaker pre-filled with 800 mL H₂O while constantly stirring with a m....

Results

Figure 1: Detailed information on the preparation of horizontal hippocampal brain slices. (A) Image of tools required for dissection and slicing of the rodent brain: (a) ±2 cm long and ±0.5 cm wide straps of filter paper (e.g., grade 413); .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Carbogen tank	Air Liquide	Alphagaz mix B50	Gasmixture CO2/O2: 5/95, purity 5
Culture dish (35 mm)	Corning Life Sciences	353001	https://ecatalog.corning.com/life-sciences/b2c/US/en/Cell-Culture/Cell-Culture-Vessels/Dishes%2C-Culture/Falcon®-Cell-Culture-Dishes/p/353001
Culture dish (90 mm)	Thermo Fisher Scientific	101VR20	https://www.thermofisher.com/order/catalog/product/101R20#/101R20
Curved forceps	Fine Science tools	11270-20	https://www.finescience.de/de-DE/Products/Forceps-Hemostats/Dumont-Forceps/Dumont-7b-Forceps/11270-20
D-(+)-Glucose monohydrate	Sigma Aldrich	16301	https://www.sigmaaldrich.com/catalog/product/sial/16301?lang=en&region=BE
Filter paper	VWR	516-0818	grade 413
Fine brush	Raphael Kaerell	8204	Size #1
Loctite 406	Henkel Adhesive technologies	Loctite 406	Super glue
Potassium chlorid	Chem-lab	CL00.1133	https://www.chem-lab.be/#/en-gb/prod/1393528
Potassium dihydrogen phosphate	Merck	104873	https://www.merckmillipore.com/BE/en/product/Potassium-dihydrogen-phosphate,MDA_CHEM-104873?ReferrerURL=https%3A%2F%2Fwww.google.com%2F
Razor blade to prepare hemispheres	SPI supplies	Safety Cartridge Dispenser - Pkg/10	GEM Scientific Single Edge Razor Blades
Razor blade for vibratome	Ted Pella Inc	121-6	double edge breakable style razor blades (PTFE-coated stainless steel)
Recovery chamber	home made - Generic	N/A	to collect and store brain slices in (see details in manuscript)
Scissors	Any company	N/A	Blade should be well sharpened and at least 15 cm long for easy decapitation
Sodium dihydrogen phosphate dihydrate	Merck	106342	https://www.merckmillipore.com/BE/en/product/Sodium-dihydrogen-phosphate-dihydrate,MDA_CHEM-106342?ReferrerURL=https%3A%2F%2Fwww.google.com%2F
Sodium hydrogen carbonate	Alfa Aesar	14707	https://www.alfa.com/en/catalog/014707/
Sodium chlorid	Fisher Scientific	S/3160/60	https://www.fishersci.co.uk/shop/products/sodium-chloride-certified-ar-analysis-meets-analytical-specification-ph-eur/10428420
Spatula	Sigma Aldrich	S9147-12EA	https://www.sigmaaldrich.com/catalog/product/sigma/s9147?lang=en&region=BE
Sucrose	VWR International Ltd.	102745C	https://es.vwr-cmd.com/ex/downloads/magazine/lupc_userguide_uk.pdf
Tubing for carbogen, perfusion and suction lines 1	Warner Instruments	64-0167	Tygon tubing (TY-50) for standard valve systems
Tubing for carbogen, perfusion and suction lines 2	Fisher Scientific	800/100/200 & 800/100/280	Smiths Medical Portex Fine Bore LDPE Tubing
Vibratome	Leica	14912000001	Semi-automatic vibrating blade microomei VT1200
Water bath	Memmert	WNB 7	https://www.memmert.be/wp-content/uploads/2019/09/Memmert-Waterbath-WNB-7.en_.pdf
Water purification system	Merck	Synergy millipore	to obtain highly purified water