Hy Everybody, I'm going mad with sealing HT-22 cells in patch clamp experiments.
Cells were seaded in DMEM with 10% FBS, 1x Glutamax and penstrep on polyLlysine coated coverslips.
- I've tried with different electrode resistances from 4 to 12 M ohms.
- extracelluar solution: NaCl 120,KCl 4.2,HEPES 10, CaCl2 2,MgCl2 0.3, Glucose 10, Sucrose 21- ph 7.4 298mOsm
- intracellular solution(mM) K-Gluconate 125,CaCl2 2, MgCl2 2,EGTA 10, HEPES 10, Na2-ATP 2,Na2-GTP 0.5 ph 7.35 295 mOsm.
Ht-22 cells are always very grouped even if they are at 50% of confluence. their membranes appear very delicates and they are likely to be broken. Any suggestion?
Very thanks Dr Moorthy for your answer..congratulation for your interesting work.
..so what is your suggestion in order to ameliorate the sealing with HT22 cell line in patch clamp experiment?
Best Regards
Marcello
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