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I study Ca voltage gated channels in DRG.
I use TEA-Cl in my external solution, but I have a problem with the stability of the cells.
Any suggestions?
Thanks
I also study VGCC in DRG. I have no that problem. my extrasolution is TEA-Cl 140; CaCl2 2; MgCl2 1.5; HEPES 10; Glu 8
Thanks for reply!
But I have another question. What is better use CaCl or BaCl?
Thanks!
I never use Ba to replace Ca in my experiment..I think it depends on your popuse of your experiment. I work on DRG neurons from 7 day old rats. I focus on intra ca concentration. In my experiment the extrasolution I gave you works well. my highest record of succeed patch is 58 cells for one slice. So maybe it's the cell culture problem..I also use aldult rat DRG neurons. I could share my protocal. tell me your E-mail..
If you have effective patch protocols and solution recipies please share them with the whole community by posting them here
http://www.scientistsolutions.com/science-protocols.aspx?ssipg=AddProtocol
The choice between Ca and Ba depends on whether you want to eliminate downstream consequences of calcium inlflux (e.g. Ca activated Ca release) during your experiment or not.
Hi!
I work on DRG neurons from 8-20 days old but not in slice with cells isolated.
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external solution:130 TEACl, 10 BaCl2, 10 glucosio, 10 HEPES, 1 MgCl, 0,001 TTX
internal solution:
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140 CsCl, 4 MgCl, 10 HEPES, 10 EGTA, 2 Na2ATP.
Thanks!
Have you tried establishing your patch in a Na+ and Ca2+ containing solution and then perfusing in the TEA Ba2+ constaining solution only after you have a stable whole cell patch?
so for example establish a stable whole cell patch in this external solution (concentrations in mM):
NaCl 145, KCl 5, CaCl2 2, MgCl2 1, and HEPES 10 (pH 7.4; NaOH)
then record Ba2+ currents in (concentrations in mM):
Tetra-Ethyl-Ammonium (TEA)-chloride 160, HEPES 10, EGTA 0.1, and BaCl2 5 (pH 7.4; TEA-OH)
the pipette solution would be (concs in mM):
Cs-methanesulfonate 103, MgCl2 4, HEPES 9, EGTA 9, (Mg)adenosine triphosphate 4, (tris)guanosine triphosphate 1, and (tris)phosphocreatine 14 (pH 7.4; CsOH)
Hi,
Does anybody know about the increase of the current after perfusion with TEA 1 mM (after perfusion with 0,1 mM), in Whole cell of Adipose derived Mesenchymal Stem cells?
I have found this effect in 5 Control cells, but none of the experimental cells
Regards,