Perforated Patch clamp with Gramicidin D

3 posts / 0 new
Last post
JunEphys
JunEphys's picture
Perforated Patch clamp with Gramicidin D

Hi,

I have a question about  How to tell if cell membrane is reptured in Perforated Patch clamping with Gramicidin D. I read that membrane rupture would result in a small depolarization in current clamp as small as 2mV but I don't know how one can notice that and be sure about the membrane rupture. I know that by measuring the I-O curve to monitor the reversal potential of Cl, one can determine if Cl movement is disrupted as well but I wonder if there's another way to tell. 

Thank you, 

Jun

TMBGJO
TMBGJO's picture
 Hey Jun,

 Hey Jun,

I add a high concentration of Ca2+ to my pipette solution, as the gramicidin pores would not let the Ca2+-ions pass. If your access into the cell would be because of membrane rupture the cell would die as a consequence of the Ca2+ inflow. Hope that helps.

TMB

The FFM
The FFM's picture
 this paper should help