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Confocal Imaging of Single Mitochondrial Superoxide Flashes in Intact Heart or In Vivo
Published on: November 5, 2013
Methionine-Derived Fluorescent Probes Targeting Mitochondria: A Tool for Real-Time Oxidative Stress Monitoring in
Niharika Pareek1, Nripankar Kalita1, Roopam Pandey1
1Molecular Sensors and Therapeutics (MST) Research Laboratory, Department of Chemistry, School of Natural Sciences, Shiv Nadar Institution of Eminence Deemed to be University, Delhi NCR, Greater Noida, Uttar Pradesh, 201314, India.
Abstract:
Reactive oxygen species (ROS) play crucial roles in both cell signaling and defense mechanisms. Hypochlorous acid (HOCl), a strong oxidant, aids the immune response by damaging pathogens. In this study, we developed two pyridinium-based fluorophores PSSM and PSSE for selective hypochlorite detection. Out of these two fluorescent probes, PSSM shows a strong turn-on emission via a photoinduced electron transfer (PeT) mechanism, excellent mitochondrial localization, and rapid response to HOCl with high selectivity among reactive oxygen species by achieving a detection limit of 2.41 μM. It successfully detects both exogenous and endogenous HOCl in live cells, enabling the study of HOCl's role at the organelle level. Structural analysis of PSSM via thioether oxidation confirmed by HPLC, NMR and HRMS further supports its specificity. Confocal imaging and flow cytometry studies further highlights its utility in investigating oxidative stress, positioning this fluorophore as a valuable tool for monitoring HOCl imbalances in biological systems.
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