Wov! seems to be a difficult task to accomplish... You patch blind and
with perforated at the same time!!!
I work also in the cortex (prefrontal cortex), and I had a run-down
problem so I decide to do perforated patch-clamp, but I use joung
animals (12-20 postanal ), so I don't have to do it blind! Is it really
so difficult as I imagine?
We have here in my laboratory some experience in striatum and cortex. If
you need some help, just let me know!
bilz0r at hotmail.com wrote:
> Thanks for replying.
>> I'm probably going to be patching pyramdal cells in the cortex (blind),
> or Nigral cells (DIC visualization) in mouse brain slices.
>>>> SJM Guzman wrote:
>>>>I have some experience with perforated patch-clamp. For Nystatin you
>>should simply to take care to work in darkness, when possible (it is
>>UV-sensitive...). Stock solution should be stored (-20ºC) in those color
>>eppendorf tubes or wrapped with Aluminum foil, to keep them away from
>>the UV rays....
>>>>I could give you more advices, if you tell me exactly which kind of
>>cells are you trying to patch (slices or cell-culture?).
>>>>I personally find Anphotericin B better for perforated patching! :P
>>>>>>bilz0r at hotmail.com wrote:
>>>>>>I've recently got in some Fluka nystatin from sigma:
>>>http://www.sigmaaldrich.com/catalog/search/ProductDetail/FLUKA/74721>>>>>>..for perforated patching. Now while I understand how to use nystatin
>>>in theory, the actual practicalities escape me. Specifically, it says
>>>on the side of the bottle, that it is UV sensitive, and that I need to
>>>store it under argon! I don't think there is any argon within a mile
>>>from my lab, what am I supposed to do when I make up my portion of DMSO
>>>stock? Or is it one of those kinda "in a perfect world, you'd store it
>>>under argon, but in reality, no one actually does this" kinda warnings?