how to deliver compound to cell for hERG assay

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zhangxu04
zhangxu04's picture
how to deliver compound to cell for hERG assay

I will make the hERG assay with patch clamp.  but I am not sure how to deliver the compound.
one way is to use the gravity perfusion, but I think this need much compound and much time for the compound reaching the cell fully
second way is to use the pressurized perfusion system ( "puff"). I think this method may affect the whole cell seal
I hope someone could make a brief comparation of these two delivering method. 
thank you very much

The FFM
The FFM's picture
most people just perfuse the

most people just perfuse the compound in to the chamber by gravity flow or another variation.
 
use as small a chamber as possible if you are worried about equilibration time.  Most people are  interested in the steady-state bock so this normally is not an issue.
 
you could make a perfusion system with a micropipette to continuously perfuse the cell but using less drug with the drug locally applied on the cell being patched rather than the whole bath.  However, a picosprizter puff will not be enough volume for block to reach equilibrium in most cases as it takes minutes for many compounds.

zhangxu04
zhangxu04's picture
thank you very much for your

thank you very much for your answer.
this issue  trouble me much.  I will choose one of these two system, if possible could you give me a brief comment :
VC³-4PG  or  VC³-8SP  all from ALA company