Slice preparation
Nicola Biagio Mercuri
Disclaimer
The protocols.io team notes that research involving animals and humans must be conducted according to internationally-accepted standards and should always have prior approval from an Institutional Ethics Committee or Board.
Abstract
Slice preparation for electrophysiology
Steps
Animal sacrifice and surgical procedure
Anesthetize the animal under halothane
Decapitate the animal with a guillotine
Gently open the bones of the skull and remove the brain, dropping it into a beaker containing ice-cold ACSF continuously bubbled with an O2/CO2 (95/5%) gas mixture
ACSF composition (in mM): 126 NaCl, 2.5 KCl,1.2 MgCl2, 1.2 NaH2PO4, 2.4 CaCl2, 10 glucose and 25 NaHCO3.
Brain slices cutting
After 1 min, place the brain on a petri dish with filter paper soaked with ACSF, and then separate the region of interest with a razor blade
Stick the peace of brain on the appropriate magnetic surface provided by the Leica VT1200S microtome with cyanoacrylate glue, and place it in the cutting chamber filled with ice-cold ACSF
Proceed with the slicing procedure (250 μm thick) and remove the surrounding tissue in each slice, to isolate the area of interest
The slices are then left to recover for at least 1hr in saturated ACSF at 32°C
