I am trying to do some perforated patch recordings using amphotericin B. It has been nearly month now and I can't have even one sucessful experiement with really stresses me out :(.
So I dissolve amphoerticin B ( powder from Sigma) in DMSO , sonicate them then add it to the normal intra and I keep this in ice all the time.
I backfill the pipette with normal saline then with intra+ amphotericin B.
I do my best to do all this in darkness as amphoericin is senstitve to light.
I noticed that the spikes of the cells increase in the begining after forming the seal then it decrease again and usually I loose the cell.
Any tips that can help me ?
Thanks in advance ...