On Tuesday 13 June 2006 18:38, jai miss wrote:
> Is there anybody who has some experience in giant patch technique
> especially in oocytes? I have some trouble with oocytes and am looking for
> the solution.
I assume that you want to record Ion channels. Not single channels but
reasonable populations. I know that it is possible to get currents of up to
several hundred picamperes from a patch. But...... with pipettes with large
diameters (we used resistances of less tha a Megaohm it takes up to 15
minutes to get a Gigaohm seal (if you get a seal). This depends on the
Oocytes, but we never found a way to predict their properties. If you get a
seal and you have good expression - let's say 100 miroamperes in two
electrode Voltage clamp (no problem with good RNA and the right channels)-
you may not have any current i the patch. Most likely because the channels
are clustered and not homogeneously distributed on the Oocyte surface. If you
than need to excise your patch you may often loose the patch after a short
while (seconds to minutes). If you achieve a success rate of 10 to 20 per
cent be happy. We tried hard, but we never found a working solution except
for trying trying trying..... So if you see an alternative (cell lines, two
electrode clamp .....) it may be a good idea to give it a chance.
Best wishes
Michael
--
Dr. Michael Ferber
Herchenbachstr. 13
D-34253 Lohfelden
Tel:(+49) 0561 8165785
mobil: (+49) 0160 96734843
email: Michael.Ferber at gmx.de
