Fast desensitizing ion channels recorded on Qube 384

雑誌名: Sophion application report
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著者: Daniel Sauter et Al.

  • The nicotinic acetylcholine receptor alpha1 (nAChR α1) is a fast desensitizing channel that makes the development of a stable high throughput assay on automated patch clamp systems challenging.
  • The solution stacking feature on Qube reduces ligand exposure times below 1 s. Using this feature, it is now possible to evoke stable acetylcholine signals for agonist concentrations at least up to EC50.
  • An assay for nAChR α1 endogenously expressed in TE671 cells with 5 µM acetylcholine showed the following characteristics:
    • Overall success rate of 89%
    • Overall whole-cell seal resistance = 1.2 ± 0.4 GΩ (SD)
    • Coefficient of variance for rows and columns in DMSO control ≤12%
    • IC50(max/min)(tetracaine) of a non-cumulative concentration response experiment = 1.5
    • Z’ greater than 0.70
  • The assay stability is also owed to unique architecture of the QChip where microfluidic channels ensure rapid and complete solution exchange of compound and agonist.