My cells break before reaching sealing resistance. its passage no. is 10. I use 5ml syringe for sealing cells. Earlier it was working fine, now a days its a daily problem. Can some body suggest reason and advice for it?
Hello Abhilasha,
You have disclosed very little information on what is your approach of getting giga-seal which makes it hard to trouble-shoot. In general, it sounds like that premature rupturing is related to excessive negative pressure you apply to pipette interior. HEK-293 are usually very easy to seal: they require slight positive pressure
Was there a change in the osmolarity of any of your solutions: culture media, recording solution and intracellular solution?
The shape and cell viability are affected by the osmolarity of the solutions used.
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