How to record cardiomyocyte action potential at 37 degree C

1 post / 0 new
findanswer3513
findanswer3513's picture
How to record cardiomyocyte action potential at 37 degree C

Dear Sir / Madam,

I can't  patch on cardiomyocytes to record action potentials at 37 degree C using a perforated patch technique. I tried many things, but failed.

Following the suggestions from this forum, I patched the cell at room temperature. After cell got gigaohm sealed, I turned on the temperature controller. At this moment, I found that  it was very easy to loss the cell, because of an electric shock?.

So late on, I made a little bit changes and did as followings: (1) I turned on the temperature controller but set temperature at 22oC; (2) patched the cell to get gigaohm seal; (3) waited for about 40 minutes to allow Gramicidin to perforate the cell membrane; (4) switched voltage clmap mode to current clamp mode to see if action potential could be induced or not and (5) if the action potential could be seen, then turned the wheel of temperature controller to increase the bath temperature to 37oC at once. But I found that the cell still losed.
 
Could any body give me some suggestions about how to record the cardiomyocyte action potential at 37oC using a perforated patch clamp technique? Should I increase the bath temperature slowly? If so, how much slow? 

Many thanks,
 
Shimin