9
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: not found

      Methods for the detection of reactive oxygen species

      Read this article at

      ScienceOpenPublisher
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          The combined usage of more than one method is recommended for better detection of reactive oxygen species.

          Abstract

          There are many pieces of evidence suggesting the mediating role of reactive oxygen species (ROS) in cell life, stress and death, and it is known that ROS at different concentrations would play distinctly different roles therein, thus eventually leading to the different fates of cells. Therefore it is highly necessary to carry out a reliable measurement of the concentration or relative level of ROS. Many means are available for ROS detection, such as the classical and commonly used spectrophotometry methods, use of fluorescence and chemiluminescence probes, and electron spin resonance (ESR/EPR), which provide direct identification of different types of oxygen radicals. In addition, fluorescent protein-based redox indicators designed for monitoring redox status changes would also be introduced in the text. Considering the fact that each type of method mentioned above has drawbacks during application, the present review tries to make a comparison and summary of their advantages and disadvantages systemically, and as a conclusion, the rational use of more than one method is recommended for better testing of the free radicals in cells.

          Related collections

          Most cited references112

          • Record: found
          • Abstract: found
          • Article: not found

          ROS as signalling molecules: mechanisms that generate specificity in ROS homeostasis.

          Reactive oxygen species (ROS) have been shown to be toxic but also function as signalling molecules. This biological paradox underlies mechanisms that are important for the integrity and fitness of living organisms and their ageing. The pathways that regulate ROS homeostasis are crucial for mitigating the toxicity of ROS and provide strong evidence about specificity in ROS signalling. By taking advantage of the chemistry of ROS, highly specific mechanisms have evolved that form the basis of oxidant scavenging and ROS signalling systems.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Fluorescence probes used for detection of reactive oxygen species.

            Endogenously produced pro-oxidant reactive species are essential to life, being involved in several biological functions. However, when overproduced (e.g. due to exogenous stimulation), or when the levels of antioxidants become severely depleted, these reactive species become highly harmful, causing oxidative stress through the oxidation of biomolecules, leading to cellular damage that may become irreversible and cause cell death. The scientific research in the field of reactive oxygen species (ROS) associated biological functions and/or deleterious effects is continuously requiring new sensitive and specific tools in order to enable a deeper insight on its action mechanisms. However, reactive species present some characteristics that make them difficult to detect, namely their very short lifetime and the variety of antioxidants existing in vivo, capable of capturing these reactive species. It is, therefore, essential to develop methodologies capable of overcoming this type of obstacles. Fluorescent probes are excellent sensors of ROS due to their high sensitivity, simplicity in data collection, and high spatial resolution in microscopic imaging techniques. Hence, the main goal of the present paper is to review the fluorescence methodologies that have been used for detecting ROS in biological and non-biological media.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Imaging dynamic redox changes in mammalian cells with green fluorescent protein indicators.

              Changes in the redox equilibrium of cells influence a host of cell functions. Alterations in the redox equilibrium are precipitated by changing either the glutathione/glutathione-disulfide ratio (GSH/GSSG) and/or the reduced/oxidized thioredoxin ratio. Redox-sensitive green fluorescent proteins (GFP) allow real time visualization of the oxidation state of the indicator. Ratios of fluorescence from excitation at 400 and 490 nm indicate the extent of oxidation and thus the redox potential while canceling out the amount of indicator and the absolute optical sensitivity. Because the indicator is genetically encoded, it can be targeted to specific proteins or organelles of interest and expressed in a wide variety of cells and organisms. We evaluated roGFP1 (GFP with mutations C48S, S147C, and Q204C) and roGFP2 (the same plus S65T) with physiologically or toxicologically relevant oxidants both in vitro and in living mammalian cells. Furthermore, we investigated the response of the redox probes under physiological redox changes during superoxide bursts in macrophage cells, hyperoxic and hypoxic conditions, and in responses to H(2)O(2)-stimulating agents, e.g. epidermal growth factor and lysophosphatidic acid.
                Bookmark

                Author and article information

                Journal
                AMNECT
                Analytical Methods
                Anal. Methods
                Royal Society of Chemistry (RSC)
                1759-9660
                1759-9679
                October 4 2018
                2018
                : 10
                : 38
                : 4625-4638
                Affiliations
                [1 ]National Reference Laboratory of Veterinary Drug Residues (HZAU)
                [2 ]MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods
                [3 ]Huazhong Agricultural University
                [4 ]Wuhan
                [5 ]China
                Article
                10.1039/C8AY01339J
                674b1ebf-3228-48ff-83d4-5bc49a439448
                © 2018

                http://rsc.li/journals-terms-of-use

                History

                Comments

                Comment on this article