TEA-Cl

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Querol
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TEA-Cl

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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

ZHANG Xuan
ZHANG Xuan's picture
I also study VGCC in DRG. I

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

Querol
Querol's picture
Thanks for reply!

Thanks for reply!

But I have another question. What is better use CaCl or BaCl?
Thanks!

ZHANG Xuan
ZHANG Xuan's picture
I never use Ba to replace Ca

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..

The FFM
The FFM's picture
If you have effective patch

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.

Querol
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Hi!

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!

The FFM
The FFM's picture
 

 
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)
 

CHES
CHES's picture
Hi,

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,