Measuring reactive oxygen and nitrogen species with fluorescent probes: challenges and limitations.
The purpose of this position paper is to present a critical analysis of the challenges and limitations of the most widely used fluorescent probes for detecting and measuring reactive oxygen and nitrogen species. Where feasible, we have made recommendations for the use of alternate probes and appropriate analytical techniques that measure the specific products formed from the reactions between fluorescent probes and reactive oxygen and nitrogen species. We have proposed guidelines that will help present and future researchers with regard to the optimal use of selected fluorescent probes and interpretation of results.
Department of Biophysics and Free Radical Research Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA. balarama@mcw.edu
Balaraman
B
Kalyanaraman
Victor
V
Darley-Usmar
Kelvin J A
KJ
Davies
Phyllis A
PA
Dennery
Henry Jay
HJ
Forman
Matthew B
MB
Grisham
Giovanni E
GE
Mann
Kevin
K
Moore
L Jackson
LJ
Roberts
Harry
H
Ischiropoulos
D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000032:analysis; D010084:Oxidation-Reduction; D030421:Peroxynitrous Acid / Q000032:analysis; D026361:Reactive Nitrogen Species / Q000032:analysis*; D017382:Reactive Oxygen Species / Q000032:analysis*; D013050:Spectrometry, Fluorescence; D013194:Staining and Labeling / Q000379:methods*; D013481:Superoxides / Q000032:analysis
D005456:Fluorescent Dyes; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; D013481:Superoxides; D030421:Peroxynitrous Acid; D006861:Hydrogen Peroxide