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

tope effect values for spontaneous oxidation ranged from 3.7-4.7, whereas values for superoxide oxidation were between 2.5-2.8 for many different types of deuterium-containing fluorescent dyes, including DHE.

"This was the key experiment that demonstrated that there was a much larger difference in the way the hydrogen and deuterium compounds reacted to spontaneous oxidation than how they dealt with oxidation by a reactive oxygen species," explained Murthy.

Murthy's collaborators W. Robert Taylor and Sarah Knight tested the ability of both types of dyes to detect reactive oxygen species inside cells. Since the deuterium-containing probes were less affected by air and light and background fluorescence was suppressed, the researchers found that the dyes more accurately detected small amounts of reactive oxygen species. Knight is an Emory University postdoctoral fellow and Taylor is a professor in the Coulter Department, the director of Emory's Division of Cardiology, and a member of the Atlanta VA Medical Center's Division of Cardiology.

Following the cellular experiments, Knight and Coulter Department postdoctoral fellow Seungjun Lee investigated whether the kinetic isotope effect would similarly improve the ability of H-Cy7 -- a hydrocyanine dye developed by Murthy -- to detect radical oxidants in vivo. In experiments, the deuterium-containing version of Cy7 generated a 10-fold difference in fluorescence intensity versus control probes compared to only a five-fold difference for the hydrogen probe.

"This new process of replacing hydrogen with deuterium is potentially valuable because the positive results are universal among many different types and classes of probes," explained Murthy. "All of the modified probes generated less background fluorescence, while maintaining high reactivity with reactive oxygen species and generating similar levels of fluorescence in cells and animals stimulated to produce them."

Murthy indica
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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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