Bacterial Biosensor Assay for Detecting Bioavailable Mercury Under Anaerobic Conditions

Samenvatting

Source: Stenzler, B. R. et. al., An Anaerobic Biosensor Assay for the Detection of Mercury and Cadmium. J. Vis. Exp. (2018)

This video demonstrates the use of genetically engineered bacterial biosensors to accurately measure the bioavailable fraction of mercury ions under anaerobic conditions. By varying the concentration of magnesium sulfate, the assay ensures that the biosensor selectively responds to free mercury ions, as indicated by changes in fluorescence intensity.

Protocol

1. Exposure Assay

1. Designing a plate layout.
   NOTE: Be sure to have all pipetting values calculated and stocks prepared prior to starting the exposure assay. How to properly design an experiment and what controls to include is detailed in the text. In addition, the experiment should not be started if there is oxygen in the anaerobic chamber indicated on the anaerobic monitor.
2. Design the plate layout according to a 96-well template. This will allow for 32 different treatments to run experiments in technical replicates of 3, which is best represented by a 4 x 8 grid to set up vials (see....

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
7 ml standard vial, rounded interior	Delta Scientific	200-007-20	34 recommended
Vial tray, 21 mm openings	Delta Scientific	730-2001	4 for (4 x 8 grid)
24 mm closure	Delta Scientific	600-024-01	32 recommended
Corning 96- well clear-bottom nonbinding surface microplate	Corning	Corning 3651
Anaerobic Chamber (glovebox)			The air in the anaerobic chamber should be (97 % nitrogen ± 2 % and 3 % hydrogen ± 2 %)
Microplate reader
450 (±10) nM filter for the microplate reader
500 (±10) nM filter for the microplate reader