When performing patch-clamp recordings on HT22 cells, I encountered technical challenges. Following enzymatic dissociation, the cells often appear shrunken, making it difficult to assess their viability. Conversely, attempting patch-clamp on adherent HT22 cells proves problematic, as achieving a high-quality seal is difficult—the seal resistance frequently remains around 50 MΩ and fails to improve further. How might this issue be addressed?
Hi Chongyi Yan,
Enzymatic dissociation may be one issue. Are you using the lowest concentration of enzyme possible? Also, which media are you using to keep the cells? Culture media have a higher osmolarity and must be washed thoroughly before any patch attempt because of the proteins in it - they make the seal formation really difficult. And your external solution for patching, does it have a reasonable osmolarity (285-315 mOsm)?
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