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The use of fluorophore‐containing spin traps as potential probes to localize free radicals in cells with fluorescence imaging methods
Author(s) -
Pou Sovitj,
Bhan Anila,
Bhadti Vishweshwar S.,
Wu Samuel Y.,
Hosmane Ramachandra S.,
Rosen Gerald M.
Publication year - 1995
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.9.11.7649408
Subject(s) - nitrone , fluorophore , nitroxide mediated radical polymerization , radical , fluorescence , chemistry , photochemistry , spin trapping , electron paramagnetic resonance , fluorescence spectroscopy , nitroxyl , spin probe , biophysics , nuclear magnetic resonance , biochemistry , organic chemistry , catalysis , radical polymerization , polymer , biology , physics , quantum mechanics , membrane , cycloaddition , copolymer
Central to the study of free radical processes is the ability to identify and localize their cellular site of formation. Under the best of experimental conditions, spin trapping/ESR spectroscopy can only characterize intracellular production of specific free radicals and confocal microsocopy can only localize the site of their formation. In this article, we report on the development of a fluorophore‐containing nitrone, α‐[4‐[5‐((2‐carboxy)phenyl)‐5‐hydroxy‐4‐oxo‐3‐phenyl)‐2‐pyrroHn‐1 ‐y l ]pheny l ]‐N‐(lert‐butyl)nitrone sodium salt (4).3 This nitrone (4) reacts with α‐hydroxyethyl radical with a second order rate constant of 1.7 X 105 M 1 s‐1 to give a characteristic ESR spectrum. However, we were unable to decrease the fluorescence emission, due in part to the small concentration of nitroxide generated from the reaction of α‐hydroxyethyl radical with nitrone (4). Using the fluorophore‐containing nitroxide (7) as a model, we found that only 12% of the nitroxide needs to be reduced to give an almost 400% increase in the fluorescent emission of (7). Our findings suggest new approaches to the development of various fluorophore‐containing nitrones that can both characterize specific free radicals and localize their site of intracellular formation.—Pou, S., Bhan, A., Bhadti, V. S., Wu, S. Y., Hosmane, R. S., Rosen, G. M. The use of fluorophore‐containing spin traps as potential probes to localize free radicals in cells with fluorescence imaging methods. FASEB J. 9, 1085‐1090 (1995)