Method Article

Obtaining Horizontal Mouse Hippocampal Brain Slices

July 8th, 2025

In This Article

Abstract

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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

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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....

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Results

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Brain slicing process; diagrams, tools, setup; specimen orientation; MRI preparation; scientific study.
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); .......

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Disclosures

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No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Carbogen tankAir LiquideAlphagaz mix B50Gasmixture CO2/O2: 5/95, purity 5
Culture dish (35 mm)Corning Life Sciences353001https://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 Scientific101VR20https://www.thermofisher.com/order/catalog/product/101R20#/101R20
Curved forcepsFine Science tools11270-20https://www.finescience.de/de-DE/Products/Forceps-Hemostats/Dumont-Forceps/Dumont-7b-Forceps/11270-20
D-(+)-Glucose monohydrateSigma Aldrich16301https://www.sigmaaldrich.com/catalog/product/sial/16301?lang=en&region=BE
Filter paperVWR516-0818grade 413
Fine brushRaphael Kaerell8204Size #1
Loctite 406Henkel Adhesive technologiesLoctite 406Super glue
Potassium chloridChem-labCL00.1133https://www.chem-lab.be/#/en-gb/prod/1393528
Potassium dihydrogen phosphateMerck104873https://www.merckmillipore.com/BE/en/product/Potassium-dihydrogen-phosphate,MDA_CHEM-104873?ReferrerURL=https%3A%2F%2Fwww.google.com%2F
Razor blade to prepare hemispheresSPI suppliesSafety Cartridge Dispenser - Pkg/10GEM Scientific Single Edge Razor Blades
Razor blade for vibratomeTed Pella Inc121-6double edge breakable style razor blades (PTFE-coated stainless steel)
Recovery chamberhome made - GenericN/Ato collect and store brain slices in (see details in manuscript)
ScissorsAny companyN/ABlade should be well sharpened and at least 15 cm long for easy decapitation
Sodium dihydrogen phosphate dihydrateMerck106342https://www.merckmillipore.com/BE/en/product/Sodium-dihydrogen-phosphate-dihydrate,MDA_CHEM-106342?ReferrerURL=https%3A%2F%2Fwww.google.com%2F
Sodium hydrogen carbonateAlfa Aesar14707https://www.alfa.com/en/catalog/014707/
Sodium chloridFisher ScientificS/3160/60https://www.fishersci.co.uk/shop/products/sodium-chloride-certified-ar-analysis-meets-analytical-specification-ph-eur/10428420
SpatulaSigma AldrichS9147-12EAhttps://www.sigmaaldrich.com/catalog/product/sigma/s9147?lang=en&region=BE
SucroseVWR International Ltd.102745Chttps://es.vwr-cmd.com/ex/downloads/magazine/lupc_userguide_uk.pdf
Tubing for carbogen, perfusion and suction lines 1Warner Instruments64-0167Tygon tubing (TY-50) for standard valve systems
Tubing for carbogen, perfusion and suction lines 2Fisher Scientific800/100/200 & 800/100/280Smiths Medical Portex Fine Bore LDPE Tubing
VibratomeLeica14912000001Semi-automatic vibrating blade microomei VT1200
Water bathMemmertWNB 7https://www.memmert.be/wp-content/uploads/2019/09/Memmert-Waterbath-WNB-7.en_.pdf
Water purification systemMerckSynergy milliporeto obtain highly purified water

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Tags

Horizontal Hippocampal SlicesMouse Brain SlicingVibratome SectioningHigh Sucrose SolutionCarbogenated FluidTissue Recovery ChamberBrain Hemisphere SecuringSlice Collection MethodHippocampal Fiber PreservationACSF Recovery Solution

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