Place the brain into pre-chilled aCSF solution.Setup vibratome by filling the buffer chamber wtih cold aCSF solution, setting the desired thickness (2-500 µM for brain slices) and adjust the cutting speed to the desired setting (in the video protocol we used number 0.15 mm/min).Chill 1 L of aCSF over dry ice until PH adjusted to 7.3 with 0.5M KOH and the osmolarity adjusted with 1M sucrose. Bubble for a further 15 minutes with carbogen. Afterwards add 4.8 ml of 1M CaCl 2 and 2.6 ml of 1M MgCl 2 using a suitable pipette to give a final concentration of: Bubble aCSF with carbogen 95% O 2/5% CO 2 for 15-20 minutes to stabilize the pH to 7.3-7.4. Shake well and make sure all salts have dissolved. Top it up to with deionised H 20 to around 1.9 L, leaving enough room to add other components later. Weigh out salts using a fine balance to make up 2 L of artificial cerebrospinal fluid (aCSF) solution.
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