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Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors
Published on: February 7, 2018
Two-photon fluorescence microscopy imaging of cellular oxidative stress using profluorescent nitroxides
Hyo-Yang Ahn1, Kathryn E Fairfull-Smith, Benjamin J Morrow
1Department of Chemistry, University of Central Florida, P.O. Box 162366, Orlando, Florida 32816-2366, United States.
Journal of the American Chemical Society
|March 3, 2012
Summary
New profluorescent nitroxide probes "light up" to image oxidative stress in cells. These probes show low toxicity and high sensitivity, offering advanced cellular imaging capabilities.
Area of Science:
- Photochemistry
- Biomedical Imaging
- Chemical Biology
Background:
- Oxidative stress is implicated in various diseases.
- Current imaging techniques for oxidative stress have limitations.
Purpose of the Study:
- To synthesize and characterize novel profluorescent nitroxide probes.
- To evaluate their potential for imaging oxidative stress in biological systems.
Main Methods:
- Synthesis of nitroxide free radicals and methoxyamine analogues.
- Photophysical property analysis, including two-photon absorption (2PA).
- Cytotoxicity assays and cellular imaging studies using fluorescence microscopy.
Main Results:
- Profluorescent nitroxides effectively masked chromophore fluorescence.
- Anthracene and fluorescein nitroxides showed significant 2PA cross sections (>800 nm).
- Probes exhibited low cytotoxicity, good cellular uptake, and sensitivity to H(2)O(2)-induced damage.
Conclusions:
- Profluorescent nitroxide probes are promising tools for oxidative stress imaging.
- High fluorescence quantum yield ratios enable sensitive detection of cellular redox environments.
- Two-photon microscopy compatibility offers advantages over one-photon imaging.
