Protocol for glial Ca²⁺ imaging in C. elegans following chemical, mechanical, or optogenetic stimulation

Caenorhabditis elegans is an exceptionally transparent model to analyze calcium (Ca²⁺) signals, but available protocols for neuronal Ca2⁺ imaging may not be suitable for studying glial cells. Here, we present a detailed protocol for glial Ca²⁺ imaging in C. elegans following three different approaches including chemical, mechanical, and optogenetic stimulation. We also provide the details for imaging analysis using Image-J. For complete details on the use and execution of this protocol, please refer to Duan et al. (2020).