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Protocol for assessing lysosomal ion channel function in mammalian cells using lysosomal patch-clamp technique

Preprint Created on 02 Jun 2026 bioRxiv

This protocol describes manual whole-endolysosome patch-clamp recordings from pharmacologically or genetically enlarged endolysosomes in cultured mammalian cells. Steps include vesicle enlargement, fabrication and fire-polishing of high-resistance pipettes, mechanical dissection and isolation of enlarged vesicles, 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.

Wang, Y., Jan, L. Y.

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