Measuring reactive oxygen and nitrogen species with fluorescent probes: challenges and limitations

Free Radic Biol Med. 2012 Jan 1;52(1):1-6. doi: 10.1016/j.freeradbiomed.2011.09.030. Epub 2011 Oct 2.


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.

Publication types

  • Review

MeSH terms

  • Fluorescent Dyes / analysis*
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • Hydrogen Peroxide / analysis
  • Oxidation-Reduction
  • Peroxynitrous Acid / analysis
  • Reactive Nitrogen Species / analysis*
  • Reactive Oxygen Species / analysis*
  • Spectrometry, Fluorescence
  • Staining and Labeling / methods*
  • Superoxides / analysis


  • Fluorescent Dyes
  • Reactive Nitrogen Species
  • Reactive Oxygen Species
  • Superoxides
  • Peroxynitrous Acid
  • Hydrogen Peroxide