Can one prepare ahead and aliquot ACSF and keep it in -20C?
The stock is made without CaCl.2H20 & D-Glucose, that will be added before bubbling and then using. (I am using the recipe: NaCL 124Mm, KCl 3Mm, KH2PO41.25Mm, MgSO4 4Mm, NaHCO3 26Mm, Acsorbic acid 2Mm, and Adenosin 0.075Mm)
The ACSF will later be used to dissect brains for organotypic slice culture.
Thanks in advance,
Orli
Dear Orli,
The quick answer is yes you can. But why? Usually 10x ACSF is very stable in fridge for a long time (~ 6 months). I would rather make a 10x every 3-6 months than defrosting every day. Just an idea.
You should also add your Magnesium when you add your Calcium while following bubbling with carbogen, as divalent cations tend to precipitate. But you can add 10x D-glucose prior, due to high osmolarity, it will help keeping bugs out.
best wishes,
Refik
I am going to do organotypic brain slice culture of mouse, unfortunately, we don't have vibrotome for cutting slice 300-400um, i will use brain matrix mold 0.5mm replacing vibrotome, is it okay? The other thing is that is oxygenated with 95% O2 is necessary? because the O2 doesn't dissoveld in the water. what kind of culture medium is best to make the culture surviving longer. I appreciate it, if you provide me with your medium and some experience.
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