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Acute slice preparation

NK Nathan J. Klett
CA Charles N. Allen
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During their light phase (ZT 1-3 for day experiments and ZT 10-12 for night experiments), animals were removed from their housing chambers, anaesthetized with isoflurane, and decapitated. The brain was quickly removed and submerged in an ice-cold slicing solution consisting of (in mM): 111 NaCl, 26 NaHCO3, 11 dextrose, 6 Na-gluconate, 4 MgCl2, 3 KCl, 1 NaH2PO4, and 0.5 CaCl2, saturated with 95% O2, 5% CO2. The brain was blocked and 175 μm thick coronal slices were prepared with a Leica VT1000S vibrating blade microtome. Slices were incubated in slicing solution for 1-4 hours at 34 °C before recording.

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