I'm studying hippocampal pyramidal and granule cells in coronal mouse brain slices using patch-clamp. I use mixed NaCl/sucrose ACSF for cutting (ice-cold) and recovery (30 min at 35-37 °C), 0.5 liters per experiment: 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.
Saturating this solution with O2/CO2 and then chilling it usually takes about two hours, and then I have to wait another two hours before I can start patch clamping. I only have a -20 °C freezer. Can I save some time by carbogenating and freezing the sucrose-based ACSF a day or a few days before an experiment and then carbogenating the ice-cold (or even slushy) solution on the day of the experiment?
Yes, you can do that.
However 2h just bubbling is too long (it should only take 20-30 minutes). Make sure you have a bubble stone making small bubbles, and chill the solution before/while bubbling (it helps dissolve more carbogen).
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