Time-lapse live-cell imaging of pyroptosis by confocal microscopy

Pyroptosis is a type of regulated necrosis executed by gasdermin. Osmotic cell swelling and membrane perforation are the key features of pyroptosis. This protocol presents time-lapse imaging of morphological changes during pyroptosis using a confocal microscope. We describe the step-by-step ectopic expression of gasdermin, cell staining with nuclear and membrane probes, and visualization of pyroptosis by time-lapse imaging. This protocol is applicable to monitoring pyroptosis in various situations. For complete details on the use and execution of this protocol, please refer to Qin et al. (2023).1