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[Neuroscience] Re: On cell recording? What goes in the pipette?

jonesmat via neur-sci%40net.bio.net (by jonesmat At physiology.wisc.edu)
Sat Dec 23 02:06:07 EST 2006


Bill wrote:
> Hi,
>
> I want to do some on cell recording, to see if a drug modulates the
> firing rate of a spontaneously active population on neurons. What
> should I fill the electrode with? Regular aCSF will get too basic
> without CO2, 0.9% NaCl will blow away K+ etc from the extracellular
> space when I come to seal onto the cell. What about regular aCSF with
> 10mM HEPES and pHed; along the lines of 130 NaCl, 2.5 KCl, 2 CaCl2, 1
> MgCl2, 10 HEPES; 1.25 NaH 2PO4, 25 glucose; pH was adjusted using NaOH
> (an aCSF recipe I've seen used when people want to alter the pH of
> slices on the whole).
>
> Thoughts?
>
> Thanks.


Ah, just fill with your normal aCSF solution. Your concerns about pH
changes etc are all valid, the pH of you pipette won't be the same as
the extracellular pH. But remember that once you make an on-cell patch,
the cell surface area covered by that patch is really tiny, like 1% or
less of the whole cell surface. So even if your solution is normal
slice aCSF, ungasssed and with weird pH, this isn't probably going to
affect the way the cell behaves, since you're not changing the
intracellular cell pH with an on-cell electrode. You'll probably be
able to monitor the cells spike firing just fine this way. Think
simple. Simple is good.

Cheers,

Matt



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