Protocol for assessing endolysosomal chloride channel function in mammalian cells using endolysosomal patch-clamp technique.

Here, we present a protocol for assessing endolysosomal chloride channel function in mammalian cells using the endolysosomal patch-clamp technique. We describe steps for vesicle enlargement, fabrication and fire-polishing of high-resistance pipettes, and mechanical dissection and isolation of enlarged vesicles. We then detail procedures for giga-seal formation and configuration to whole-endolysosome modes. For complete details on the use and execution of this protocol, please refer to Wang et al.1.