Electroencephalography-Based Recording Technique to Record the Epileptiform Activity

Abstract

Source: Carvalho, M. et al., A Model of Epileptogenesis in Rhinal Cortex-Hippocampus Organotypic Slice Cultures. J. Vis. Exp. (2021).

This video describes the technique for recording the epileptiform activity of rhinal cortex-hippocampus organotypic slices ex vivo using electrophysiology or EEG set-up. This helps to study the dynamics and progression of epileptogenesis and screen potential therapeutic targets to treat epilepsy.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Preparation of rhinal cortex-hippocampus slices NOTE: The preparation of rhinal cortex-hippocampus slices uses P6-7 Sprague-Dawley rats. Culture setup and medium preparation On the day before the culture, prepare the required media and place them at 4 °C. Prepare d…

Representative Results

Figure 1: Detailed procedure for the preparation of rhinal cortex-hippocampus organotypic slices. (A) Remove the brain from the head and place it in ice-cold GBSS with the dorsal surface faced up. (B) Insert the forceps into the cerebellum. Open the brain through the midline and remove the excess tissue over the hippocampus. (C) With a spatu…

Materials

Artificial cerebrospinal fluid (aCSF)			Homemade
B-27™ Supplement (50X), serum free	Thermo Fisher Scientific	17504-044
Blades for scalpel handle	Fine Science Tools	10011-00
Bovine Serum Albumin (BSA)	NZYTech	MB04602	5% BSA is used to dilute the primary antibodies. Add 0.5g BSA in 10 mL PBS.
Brain/Tissue Slice Chamber System	Warner Instruments
Cell culture inserts, 30 mm, hydrophilic PTFE	Millipore SAS	PICM03050
Conventional incubator	Thermo Scientific Heraeus	BB15, Function Line	Set to 37 °C and 5% CO2
Gey’s Balanced Salt Solution (GBSS)	Biological Industries	01-919-1A
Glass Electrodes	Science Products	GB150F-10	Round tips homemade
Interface chamber	Warner Instruments	BSC-HT Haas Top
Iris Spatula Curved	Fine Science Tools	10092-12
Lens Cleaning Paper	TIFFEN
L-Glutamine solution 200 mM (Q)	Thermo Fisher Scientific	25030-024
Tissue Chopper	The Mickle Laboratory Engineering CO. LTD.	MTC/2	Set to 350 μm