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[Neuroscience] Giant patches from oocytes

Dr. Michael Ferber Michael.Ferber at gmx.de
Tue Jun 13 16:57:22 EST 2006


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



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