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Visualizing cytoplasmic ATP in C. elegans larvae using PercevalHR.

STAR protocols (2022-06-07)
Aastha Garde, David R Sherwood
RÉSUMÉ

Measuring ATP levels within the cytosol of living cells in animals is important to understand how cellular activities are energetically supported, but is challenging because of tissue complexity and ATP sensor limitations. In this protocol, we describe how to quantify ATP levels using PercevalHR in C. elegans larvae during anchor cell invasion. PercevalHR is a fluorescent biosensor that reports the cytoplasmic ATP:ADP ratio. The protocol can be adapted to analyze the ATP:ADP ratios within other cell types in C. elegans. For complete details on the use and execution of this protocol, please refer to Garde et al. (2022).

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Sigma-Aldrich
Rotenone, A mitochondrial toxin and a potent, reversible, and competitive inhibitor of complex I (NADH-CoQ reductase) of the respiratory chain.
Sigma-Aldrich
(−)-Tetramisole hydrochloride, ≥99% (GC)