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Zinc measurements with fluorescent probe FluoZin-3

RM Romaric Magerand
PR Pascal Rey
LB Laurence Blanchard
AG Arjan de Groot
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Experiments to detect free intracellular Zn2+ were based on a previously described protocol65. D. deserti was grown to OD600 0.4. Bacteria were resuspended in fresh TSB/10 and then incubated with 5 µM FluoZin‐3, AM, cell permeant (Thermo Fisher Scientific, F24195) for 60 min at room temperature in the dark to allow complete de‐esterification of intracellular acetoxymethyl (AM) esters. The bacteria were then washed three times in TSB/10. Fluorescence measurements were performed in a microplate at 28 °C and using a spectrofluorimeter (Infinite 200 Pro; Tecan). After excitation of the sample at 485 nm, fluorescence emission was recorded at 519 nm before and after addition of H2O2 or ZnCl2.

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