STAR Protocols (Jun 2023)

Protocol to assess metabolic activity of Pseudomonas aeruginosa by measuring heat flow using isothermal calorimetry

  • Katrin Beilharz,
  • Kasper Nørskov Kragh,
  • Blaine Fritz,
  • Julius B. Kirkegaard,
  • Tim Tolker-Nielsen,
  • Thomas Bjarnsholt,
  • Mads Lichtenberg

DOI
https://doi.org/10.1016/j.xpro.2023.102269
Journal volume & issue
Vol. 4, no. 2
p. 102269

Abstract

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Summary: Here, we present a protocol for assessing metabolic activity of bacterial populations by measuring heat flow using isothermal calorimetry. We outline the steps for preparing the different growth models of Pseudomonas aeruginosa and performing continuous metabolic activity measurements in the calScreener. We detail simple principal component analysis to differentiate between metabolic states of different populations and probabilistic logistic classification to assess resemblance to wild-type bacteria. This protocol for fine-scale metabolic measurement can aid in understanding microbial physiology.For complete details on the use and execution of this protocol, please refer to Lichtenberg et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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