We are an electrophysiology lab out of Milwaukee. Recently, due to ice buildup, a small crack in the door of our -20 freezer resulted in all of the freezers contents reaching room temperature during a weekend. The freezer was found with all the contents at slightly below room temp that Monday.
Among the freezers contents was internal solution. Since the freeze thaw we haven't been able to reliably obtain gigaohm seals. Just yesterday we were patching onto a 2.5 month old F344 rat and out of 35 attempts I could only get 2 gigaohm seals. Can freeze thaw impact interal solution such that gigaohm seals become difficult to obtain?
We make aCSF fresh every day. Our internal solution is a Kglu internal with biocytin. All help is truly appreciated. Thank you!
Very likely. Gluconate solubility doesn't respond well to temperature changes - especially rapid ones. Re-freezing should be OK if it doesn't stay for long above 0 °C (eg. 2-3 minutes, to check osmolarity). If it thawed to room temperature, however, you probably need a new one anyway -ATP/GTP is quite unstable.
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