Abstract
The NLRP3 inflammasome senses the activity of pore-forming toxins secreted by Staphylococcus aureus. The bacterial toxins compromise plasma membrane integrity which activates the NLRP3 inflammasome to induce host pore-forming proteins and cellular suicide, termed pyroptosis. Host cell death rates are routinely determined at pre-defined time points and on whole cell populations. To capture the dynamic interactions between bacterial pore-forming toxins and host cell death factors, we have applied live-cell imaging techniques capable of analyzing single cell events in real time. Here, we describe methods using live-cell imaging to determine the host responses, such as plasma membrane integrity, mitochondrial health, and apoptotic caspases, towards pore-forming toxins.
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Acknowledgment
We acknowledge the expert help from Keith E. Schulze and members of Monash Micro Imaging facilities for writing the MetaMorph journal scripts and ImageJ macro. The work in the Naderer Lab is supported by the National Health and Medical Research Council (grant number: 1163556). TN is an Australian Research Council Future Fellow (FT170100313).
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Chow, S.H., Naderer, T. (2022). Visualizing Effector Triggered Immunity in Response to Pore-Forming Toxins by Live-Cell Imaging. In: Kufer, T.A., Kaparakis-Liaskos, M. (eds) Effector-Triggered Immunity. Methods in Molecular Biology, vol 2523. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2449-4_15
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DOI: https://doi.org/10.1007/978-1-0716-2449-4_15
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