Ali Yehia
Learn how sample usage was reduced while simultaneously increasing throughput
Compound or sample availability can often be a limiting factor in drug/target investigations. This issue is of particular concern when compounds being tested are of a biological or synthetic origin. Recently, investigators of idiopathic hypersomnia, a condition characterized by excessive daytime sleepiness and previously attributed in some form to endogenic potentiation of GABAA receptor, were able to leverage the small sample requirements of the IonFlux automated patch clamp system and expand their investigations in finding a therapeutic solution. Their success in increasing throughput, while simultaneously reducing cost, is presented below.
Overview
Kaplan et al. set out to follow-up on previous results from their lab that found a bioactive component in the cerebrospinal fluid (CSF) extracted from patients with idiopathic hypersomnia. CSF, when applied with a low dose of GABA, increased receptor activity similar to a positive allosteric modulator. Flumazenil was shown to alleviate the problem. Hence, the investigators sought to determine the possibility of other common therapeutics that could potentially help with this condition.
Challenges
The team required a patch clamp system that used minimal precious sample to investigate multiple therapeutics in the same investigation.
Outcomes
The IonFlux system met the team's needs with its low sample volume requirements and 384-well plate capabilities. This enabled the team to gather more data faster than possible with manual patch clamp while preserving precious CSF.
Key Benefits Realized in the Lab
- Reduced Sample Usage: The investigators found that using IonFlux allowed for a 2-10-fold reduction in sample usage compared to manual patch clamp.
- Increased Throughput: IonFlux enabled the investigators to study 11 compounds within one investigation while collecting 2000-4000 data episodes per hour.
- Less Compound Usage: The microfluidic setup of IonFlux only required a low volume of drug (50 µL) & 90% could be reclaimed for subsequent analysis.
- Reduced Variability: The simultaneous measurement of up to 20 cells using IonFlux reduced cell-cell variability in EC50.
Collect Large Datasets
The available 384-well plate format of the IonFlux system enables investigators to collect large datasets while using minimal sample volume.
Drawbacks of Manual Patch Clamp
Although the manual patch clamp technique remains the "gold standard," high-throughput experimentation using manual patch clamp is extremely difficult.
Limitations of Manual Patch Clamp:
- Sample consumption is much higher due to the size of the recording environment
- Extensive training and a high skill level are required to successfully perform high-quality manual patch clamp
- Parallel recordings are not feasible without investment in multiple setups and personnel
IonFlux Microfluidics: The Key to Precise and Rapid Investigation of Multiple Compounds
To address these issues, IonFlux was designed using a microfluidic system, which enables either single-cell or simultaneous measurement of 20 cells with giga-ohm seal quality. Furthermore, the microplate setup of IonFlux allows investigation of up to 256 single compounds or concentrations in one assay, while allowing for continuous flow and rapid wash-out. Perhaps most exciting, due to its plate reader design and simplicity, IonFlux does not require a highly skilled technician for operation.
