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Whole Cell Patch Clamp of Dispersed Human Islet Cells v1

Cells use exocytosis to secrete a wide variety of molecules, including proteins, hormones, and neurotransmitters. Exocytosis can be monitored at the single-cell level by using patch-clamp electrophysiology to measure changes in membrane capacitance as vesicles fuse with the cell membrane and release their content. Dispersion of pancreatic islets into single cells allows for individual characterization of electrophysiological characteristics and allows for collection of cellular content for recovery of full-length transcriptomes by use of Smart-seq2. Described in this protocol is the dispersion of pancreatic islets into single cells followed by whole-cell patch clamp electrophysiology which includes parameters representing cell size, exocytosis, sodium channel currents, and calcium channel currents. Cells are then collected individually after recording to be processed for single-cell RNA sequencing.