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      Optimization of the C11-BODIPY(581/591) dye for the determination of lipid oxidation in Chlamydomonas reinhardtii by flow cytometry.

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          Abstract

          Lipid oxidation is a recognized end point for the study of oxidative stress and is an important parameter to describe the mode of micropollutant action on aquatic microorganisms. Therefore, the development of quick and reliable methodologies probing the oxidative stress and damage in living cells is highly sought. In the present proof-of-concept work, we examined the potential of the fluorescent dye C11-BODIPY(591/581) to probe lipid oxidation in the green microalga Chlamydomonas reinhardtii. C11-BODIPY(591/581) staining was combined with flow cytometry measurements to obtain multiparameter information on cellular features and oxidative stress damage within single cells. First, staining conditions were optimized by exploring the capability of the dye to stain algal cells under increasing cell and dye concentrations and different staining procedures. Then lipid oxidation in algae induced by short- and long-term exposures to the three metallic micropollutants, copper, mercury, and nanoparticulate copper oxide, and the two organic contaminants, diethyldithiocarbamate (DDC) and diuron was determined. In this work we pointed out C11-BODIPY(591/581) applicability in a wide range of exposure conditions, including studies of oxidation as a function of time and that it is suitable for in vivo measurements of lipid oxidation due to its high permeation and stability in cells and its low interference with algal autofluorescence. © 2013 International Society for Advancement of Cytometry.

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          Author and article information

          Journal
          Cytometry A
          Cytometry. Part A : the journal of the International Society for Analytical Cytology
          1552-4930
          1552-4922
          Oct 2013
          : 83
          : 10
          Affiliations
          [1 ] Environmental Biogeochemistry and Ecotoxicology, Institute F.-A. Forel, Earth and Environmental Sciences, Faculty of Sciences, University of Geneva, 10, route de Suisse, CH-1290, Versoix, Switzerland.
          Article
          10.1002/cyto.a.22338
          23943236
          Copyright © 2013 International Society for Advancement of Cytometry.

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