[Neuroscience] Re: Re-sealing... What ideas do you have about optimal patching solutions

Christian Wilms usenet02 at out-of-phase.de
Tue Jul 5 05:13:08 EST 2005

<bilz0r at hotmail.com> wrote:

> My supervisor completely believes in the dogma published in Sakmann and
> Neher's Single Channel Recording, that the best patching solution is
> hypotonic (270mM) and has high calcium chelation (10mM EGTA, 1mM
> CaCl2).

Just a thought on the side:
depending on what you are doing, high calcium chelation will mess things
up quite a bit. On one side, it distorts Ca-transients, through
buffering effects (Neher has quite a bit of publications on this) and
chelators also function as calcium shuttles, spreading a possibly highly
localised calcium-signal over quite large a region (Svoboda goes into
this in some of his work).
For those reasons, though high chelation tends to give better seals,
labs working on calcium-signaling tend to use low chelation (we use 50uM
in electrophysiology and 200uM for calcium imaging - some labs actually
use 50uM even for imaging purposes).
Even when you are not explicitly working on calcium signalling, one
shouldn't forget, that much of the cellphysiology being studied is
connected to calcium signalling (long-term plasicity and
endocannabinoids to name just a few).

Regards, Christian

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