drift

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lazy
lazy's picture
drift

I startted to work with the bath solution with physioloigcal temperature (34 - 37C). I am facing to a difficulty of drift:

The electrode slightly moves depending on a small change of temperature. +/- 1C (one degree) gives a changement of the electrode position and the current trace as the result. (I am using Sutter MP225.)

How people can get a stable current trace for a long period?

And also, how long the patch (WC) lasts in good condition in gengeral?

Fraser Moss
Fraser Moss's picture
Try patching and then lifting

Try patching and then lifting the cell off the coverslip.

The drift is probably attributable to the plasma membrane becoming more "fluid" as the temperature increases. As this happens there will be forces pulling between the cell's attachment to the coverslip at the base and between the membrane and the pipette tip at the top.

At room temp, you form a Giga seal and then gradually lift the whole cell up and off the dish then go whole cell. Then raise the temperature in the bath and now your recordings may suffer from less drift because the only anchor point is now from the patch pipette.

To record from cells still attached to coverslips your manipulators have to be absolutely drift free.

See this protocol book chapter for an example of physiological temp recordings from Megakaryocytes using the patch and lift technique.

Chapter title: Patch-Clamp Recordings of Electrophysiological Events in the Platelet and Megakaryocyte

Book Series Methods in Molecular Biology
ISSN 1064-3745 (Print) 1940-6029 (Online)
Volume 273
Book Platelets and Megakaryocytes
Publisher Humana Press

Abstract
http://www.springerlink.com/content/t11u3r288u7747r7/

Full-text if you have the appropriate access

http://www.springerlink.com/content/t11u3r288u7747r7/fulltext.html

lazy
lazy's picture
Great thanks! Frasermoss.

Great thanks! Frasermoss.

I appreciate a lot for your suggestion.