TOPIC

Fast desensitizing ion channels recorded on Qube 384

Sophion application report

Daniel Sauter et Al.

2017

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.

Go to journal