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2. How does Bioluminex work?
The Bioluminex assay is an innovative application of two existing technologies. It couples the separation power of high performance, thin-layer chromatography (HPTLC) with the biosensor properties of the bioluminescent marine organism Vibrio fischeri, a bacterium widely used for toxicity screening of water. TLC is an inexpensive and rapid technique that uses adsorption and capillary action to separate complex mixtures into discrete zones. This technique provides a characteristic distribution pattern of compounds that is dependent on sample composition. The TLC plate is then coated with the non-pathogenic bioluminescent bacteria, Vibrio fischeri, which emit light as a by-product of cellular respiration. As Vibrio fischeri cells reach a critical cellular density the bioluminescence reaction catalyst, luciferase is expressed. Luciferase, in the presence of oxygen, then catalyzes an oxidation reaction that releases excess energy in the form of light. This observed bioluminescence reflects the metabolic status of the cell and will decrease for cells exposed to toxic substances. Thus a reduction in light emission is a measure of toxicity that can be selectively viewed directly on the TLC plate.
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