[Neuroscience] Re: whole cell recording from dispersed neurons

Bill connelly.bill at gmail.com
Wed Apr 5 19:35:36 EST 2006

That shouldn't be a problem, our primary cultures are grown in
neurobasal media too and recorded from in a very similar (room
temperature) aCSF.

So they all die, as soon as you take them out of the incubator? Or do
they die only after you patch them. If it's the later, I suspect your
electrode, pipette solution, or break in technique. If it is the
former, then I have no idea. My only suggestion is to half, or quater
your calcium chloride, and drop your KCl down to 2.5mM. Oh, and try
30mM glucose.

I know they sound like silly changes, but anything is possible?

(my cell culture extracellular saline (mM) NaCl 142, KCl 2.5, CaCl,
2-0.5, MgCl 2, HEPES 5, Glucose 30. pH 7.4)

> Many thanks for your reply. By dead I mean depolarized and very
> leaky. Also, if by popped you mean the nucleus is visible, then, yes,
> they have popped. But I think the membrane has simply broken down- it
> looks ragged and very blebby within 20 - 30 minutes.

> And yes, these are cultured primary neurons (from hippocampus), grown
> on glass coverslips for 2 - 3 weeks. Perhaps importantly, they are
> not grown on beds of glia or in serum. They're grown in
> Neurobasal/B27.

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