We are analyzing https://link.springer.com/article/10.1007/s00216-011-4764-2.

Title:
Mitochondrial ROS production under cellular stress: comparison of different detection methods | Analytical and Bioanalytical Chemistry
Description:
Reactive oxygen species (ROS) are involved in the regulation of many physiological processes. However, overproduction of ROS under various cellular stresses results in cell death and organ injury and thus contributes to a broad spectrum of diseases and pathological conditions. The existence of different cellular sources for ROS and the distinct properties of individual ROS (their reactivity, lifetime, etc.) require adequate detection methods. We therefore compared different models of cellular stress and various ROS-sensitive dyes—2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA), MitoSOX™, and MitoTracker® red CM-H2XRos—using a confocal fluorescent imaging approach, which has the advantage of not only detecting but also of localizing intracellular sources for ROS. Confocal acquisition of DCF-DA fluorescence can be combined with ROS detection by the mitochondria-specific probes MitoSOX™ and MitoTracker® red CM-H2XRos. Specificity was controlled using various antioxidants such as Trolox and N-acetylcysteine. Using different fluorescent ROS-sensitive probes, we detected higher ROS production equally under cell starvation (IL-3 or serum depletion), hypoxia–reoxygenation, or treatment of cells with prooxidants. The detected increase in ROS was approximately threefold in IL-3-depleted 32D cells, approximately 3.5-fold in serum-deprived NIH cells, and 2.5-fold to threefold in hypoxic HL-1 cells, and these findings agree well with previously published spectrofluorometric measurements. In some cases, electron spin resonance (ESR) spectroscopy was used for the validation of results from confocal fluorescent imaging. Our data show that confocal fluorescent imaging and ESR data are in good agreement. Under cellular stress, mitochondrial ROS are released into the cytoplasm and may participate in many processes, but they do not escape from the cell.
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article, google, scholar, cas, ros, cell, biol, physiol, kuznetsov, cellular, research, mitochondrial, stress, kozlov, fluorescent, chem, andrey, med, innsbruck, privacy, cookies, content, data, kehrer, redl, hermann, troppmair, confocal, imaging, free, radic, res, schumacker, biochem, austria, information, publish, search, production, detection, published, ingeborg, fluorescence, cells, access, chapter, mol, surgery, log, journal,

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electron spin resonance mitochondria-specific probes mitosox™ fluorescent ros-sensitive probes confocal fluorescent imaging month download article/chapter dcf-da fluorescence mitotracker® red cm-h2xros mitotracker® red cm-h2xros de souza-pinto nc ros-sensitive dyes—2′ serum-deprived nih cells 6-tetramethylpiperidine-n-hydroxyl ros il-3-depleted 32d cells bioanalytical chemistry aims full article pdf mitochondrial ros production privacy choices/manage cookies related subjects cellular stress redox-sensitive fluorophores check access instant access european economic area martina haller hypoxic hl-1 cells glutathione redox equilibrium drosophila neuron subtypes n-acetylcystein pph millan-crow la bimpong-buta ny austrian science fund article analytical cellular stresses results localizing intracellular sources kozlov av conditions privacy policy accepting optional cookies confocal acquisition innsbruck medical university main content log article kuznetsov spectroscopy mitochondrial ros journal finder publish article log author information authors detection methods heinz redl kuznetsov av research grant

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         headline:Mitochondrial ROS production under cellular stress: comparison of different detection methods
         description:Reactive oxygen species (ROS) are involved in the regulation of many physiological processes. However, overproduction of ROS under various cellular stresses results in cell death and organ injury and thus contributes to a broad spectrum of diseases and pathological conditions. The existence of different cellular sources for ROS and the distinct properties of individual ROS (their reactivity, lifetime, etc.) require adequate detection methods. We therefore compared different models of cellular stress and various ROS-sensitive dyes—2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA), MitoSOX™, and MitoTracker® red CM-H2XRos—using a confocal fluorescent imaging approach, which has the advantage of not only detecting but also of localizing intracellular sources for ROS. Confocal acquisition of DCF-DA fluorescence can be combined with ROS detection by the mitochondria-specific probes MitoSOX™ and MitoTracker® red CM-H2XRos. Specificity was controlled using various antioxidants such as Trolox and N-acetylcysteine. Using different fluorescent ROS-sensitive probes, we detected higher ROS production equally under cell starvation (IL-3 or serum depletion), hypoxia–reoxygenation, or treatment of cells with prooxidants. The detected increase in ROS was approximately threefold in IL-3-depleted 32D cells, approximately 3.5-fold in serum-deprived NIH cells, and 2.5-fold to threefold in hypoxic HL-1 cells, and these findings agree well with previously published spectrofluorometric measurements. In some cases, electron spin resonance (ESR) spectroscopy was used for the validation of results from confocal fluorescent imaging. Our data show that confocal fluorescent imaging and ESR data are in good agreement. Under cellular stress, mitochondrial ROS are released into the cytoplasm and may participate in many processes, but they do not escape from the cell.
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            Characterization and Evaluation of Materials
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         Characterization and Evaluation of Materials
         Food Science
         Monitoring/Environmental Analysis
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