High-throughput automated patch-clamp of lysosomal channel TRPML1 using Sophion Bioscience platforms

Traditional whole-cell patch clamp of lysosomes is laborious and extremely specialized, yet the lysosome contains a myriad of ion
channel and transporter targets. Enhancing clearance of aggregates through lysosomal activation has proposed implications in neurodegenerative diseases such as Parkinson’s Disease, and more recently ion channel-specific rare genetic diseases. Sophion Bioscience
has therefore developed a method for high-throughput automated patch clamp (APC) of enlarged lysosomes (Lysopreps provided by Oria Bioscience) using the Qube 384 and QPatch 48 platforms.

This APC method involves using small-hole, high-resistant consumables (QChip Custom and QPlate HiR) for the purpose of capturing
organelles and forming tight seals, which enable stable recordings of ion channels in their endogenous environment. We have demonstrated the ability to capture lysosomes with a success rate of 50-70% using physiological pH and without the use of seal enhancers, such as fluoride in the internal solution. Further, we are able to record ion channel currents and concentration-response curves for TRPML1, suitable for high-throughput compound screening.