Optimization of the C11‐BODIPY^581/591 dye for the determination of lipid oxidation in Chlamydomonas reinhardtii by flow cytometry

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. V C 2013 International Society for Advancement of Cytometry

Mechanisms of toxic action of Ag, ZnO and CuO nanoparticles to selected ecotoxicological test organisms and mammalian cells in vitro: A comparative review

“…Lipid peroxidation caused by CuO NPs has also been demonstrated in tissue and gills of rainbow trout , bacteria E. coli and Bacillus subtilis (Applerot et al, 2012) and green algae C. reinhardtii (Cheloni & Slaveykova, 2013).…”

Effects of copper-oxide nanoparticles, dissolved copper and ultraviolet radiation on copper bioaccumulation, photosynthesis and oxidative stress in the aquatic macrophyte Elodea nuttallii

“…Before use, the stock dispersion received a 1 min sonication probe treatment at an amplitude of 100% (130 W, 20 kHz) without pulses (VCX 130,Sonics Vibra Cell,Sonic & Materials Inc.,Newtown,USA) and was diluted in the exposure medium. The exposure medium contained 10 À4 M KH 2 PO 4 ; 1.5 Â 0 À4 M K 2 HPO 4 ; 1.7 Â 10 À3 M NH 4 NO 3 ; 8.5 Â 10 À5 M CaCl 2 Á2H 2 O; 10 À4 M MgSO 4 7H 2 O in 10 À3 M 2-(N-morpholino) ethanesulfonic acid (MES, Sigma Aldrich, Buchs Switzerland) at pH 6.8 (Cheloni and Slaveykova, 2013) to a nominal NP concentration of 10 mg L À1 CuO. The evolution of the hydrodynamic size and zeta-potential in CuO-NP dispersions was determined in the exposure medium after 1, 2, 4 and 24 h by measurement with a Malvern Zetasizer Nano-ZS.…”

Methods Employed in Cytofluorometric Assessment of Eryptosis, the Suicidal Erythrocyte Death

“…This reaction is commonly named lipid peroxidation [96]. In order to quantify this membrane damage, the dye BODIPY 581/591 C11 (Thermo Fisher Scientific, MA, USA) is used [97]. This dye is a fatty acid analogue with particular fluorescent properties.…”