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Replacing hydrogen in fluorescent dyes improves detection ability, stability and shelf life
Date:7/20/2010

significant implications for companies that produce fluorescent probes and other compounds, according to Murthy, because commercializing and shipping the modified probes will be easier.

Another advantage for scientists using deuterium-containing fluorescent probes is that after reacting with reactive oxygen species, the probes produce the same fluorescent dye that their hydrogen counterparts produce.

"This is important from a practical standpoint because scientists have developed protocols with DHE and other fluorescent probes that they will be able to continue using by simply substituting the more stable and accurate deuterated version into the assay," explained Murthy.

Fluorescent probes detect reactive oxygen species by undergoing a chemical process called amine oxidation. The mechanism of amine oxidation for reactions involving reactive oxygen species differs significantly from reactions involving air and light. In addition, reactions with deuterium-containing probes occur at a much slower rate because deuterium is a heavier atom.

Murthy and Kundu decided to use these mechanistic and kinetic differences to selectively slow the oxidation of the fluorescent probes by air and light while maintaining their reactivity with cellular reactive oxygen species. To test the selective suppression of oxidation, the researchers examined the kinetic isotope effect -- a value that measures the ratio of the rate of a chemical reaction with hydrogen compared to the same reaction with deuterium to air and radical oxidation.

They investigated the ability of the deuterium-containing probes to compete with a common enzyme for superoxide -- a reactive oxygen species that is a form of molecular oxygen with one extra electron. The researchers found that the probes' oxidation mechanism with superoxide was different than for spontaneous oxidation because the two reactions exhibited different values for the kinetic isotope effect. Kinetic iso
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Contact: Abby Vogel Robinson
abby@innovate.gatech.edu
404-385-3364
Georgia Institute of Technology Research News
Source:Eurekalert  

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