Calcium signaling is an emerging mechanism by which bacteria respond to environmental cues. To measure the intracellular free-calcium concentration in bacterial cells, [Ca], a simple spectrofluorometric method based on the chemical probe Fura 2-acetoxy methyl ester (Fura 2-AM) is here presented using Pseudomonad bacterial cells. This is an alternative and quantitative method that can be completed in a short period of time with low costs, and it does not require the induction of heterologously expressed protein-based probes like Aequorin. Furthermore, it is possible to verify the properties of membrane channels involved in Ca entry from the extracellular matrix. This method is in particular valuable for measuring [Ca] in the range of 0.1-39.8 µM in small cells like those of prokaryotes.
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| http://dx.doi.org/10.21769/BioProtoc.3949 | DOI Listing |
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