Ultra-Fast Compound Application Services
In case your target is a ligand-gated channel, you will need a method with an ultra-fast compound application to study the effects of your drug.
Wash-in and wash-out within μs
Our automated patch clamp system uses a sophisticated piezo driven device that holds a theta tube for ultra-fast application (A), obtaining an optimal performance that enables compound exchange within milliseconds. Before usage, we calibrate and validate the correct positioning of the theta tube in a cell-free experiment, which consists of recording a liquid junction potential shift by perfusing pure water (B). Thus, confirming that our system achieves solution exchange rates in the range of a few hundred µs (C).
Distinction between deactiviation and desensitisation
The ultra-short exposure times enables us to distinguish between deactivation and desensitization. In this validation study, we used TE671 cells, which express α1β1γδ nicotinic acetylcholine receptors constitutively. TE671 cells were stimulated with short (2 ms; A-B) and long (1 s; C-D) pulses of 100 µM ACh. In line with the literature, we obtained deactivation and desensitization time constants of 9 and 19.1 ms, respectively.
Fast double pulses reveal recovery from desensitisation
Our system has an impressive ability to perfuse solutions ultra-fast and precisely timed. For instance, in this study, we used TE671 cells, which express α1β1γδ nicotinic acetylcholine receptors constitutively. We stimulated the cells with 1 mM ACh during 20 ms every 40 ms (A), to avoid the full desensitization of α1β1γδ nicotinic acetylcholine receptors. (B) Close-up of the onset of nACh-induced current from A. (C) Series of superimposed double applications with varying interpulse intervals showing recovery from desensitization.
- Danker T, Braun F, Silbernagl N, Guenther E. Catch and Patch: A Pipette-Based Approach for Automating Patch Clamp That Enables Cell Selection and Fast Compound Application. ASSAY and Drug Development Technologies 2016 | Contact an author.