I've recently started to patch dentate gyrus granule cells in coronal mouse brain slices (300 μm). I'm using mixed NaCl/sucrose ACSF for cutting (ice-cold) and recovery (30 min at 35-37°C) and standard ACSF for storage and recording.
Mixed NaCl/sucrose ACSF: NaH2PO4 1.25 mM, NaCl 87 mM, KCl 2.5 mM, MgCl2 7 mM, CaCl2 0.5 mM, glucose 25 mM, NaHCO3 25 mM, sucrose 75 mM. Osmolarity: 335 mOsm.
Standard ACSF: NaH2PO4 1.25 mM, NaCl 126 mM, KCl 3 mM, MgSO4 1.25 mM, CaCl2 1.25 mM, glucose 10 mM, NaHCO3 26 mM. Osmolarity: 300 mOsm.
From what I've learned reading research papers, some people seem to use the same sucrose-based ACSF, but they use it not only for cutting and recovery but also for storage. Lowering glucose to 10 mM in the sucrose-based ACSF also seems to be common.
Does anyone have experience storing brain slices in normal ACSF and sucrose-based ACSF? Is there any difference in slice quality or cell properties?
In my experience, storing in sucrose-based ACSF will produce what appear to be gorgeous slices, but most of the cells are actually dead. I used to limit recovery in sucrose to 20 min-ish. Nowadays in my lab, people cut in sucrose and go straight to recovery/storing in ACSF. We use 25 mM glucose in our ACSF. Good luck.
-Matthew