Flow cytometric assessment of unicellular Chlorella cells alterations under heavy metals exposure

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Article Title:	Flow cytometric assessment of unicellular Chlorella cells alterations under heavy metals exposure
Authors:	Constantin-Marian Petrescu, Violeta Turcus, Daniela Bratosin
Affiliation:	Faculty of Natural Sciences, Engineering and Informatics, ″Vasile Goldis″ Western University of Arad, Romania Institute of Life Sciences,″Vasile Goldis″ Western University of Arad, Romania National Institute for Biological Science Research & Development (INCDSB), Bucharest, Romania
Abstract:	Among all organisms in aquatic ecosystems, microalgae are key targets in pollution cases, for two basic reasons: their eco-physiological similarities with terrestrial plants (the potential sensitivity of the same metabolic processes) and their role as primary producers. Any change in the proliferation of the primary producers could provoke a global alteration in the equilibrium of the aquatic ecosystems. These characteristics support the use of the freshwater microalgae in laboratory toxicological assays. Flow cytometric analyses of Chlorella fusca cells were performed in a FACScan analyzer or Cytomics FC 500, both equipped with an argon-ion excitation laser (488 nm), detectors of forward (FSS) and side (SSC) light scatter and fluorescence detectors. For each analyzed parameter, data were recorded in a logarithmic scale and results were expressed as mean values obtained from histograms in arbitrary units. Fluorescence of chlorophyll a (>645 nm) was used as a FCM gate to exclude non-microalgal particles. To understand the mechanisms underlying the process of cell death by heavy metals action, algal cells were exposed to different concentrations of metals (Al and Cd) and analyzed by flow cytometry for morphological changes (light scattering properties) and microscopic analysis. Our results show that microalgae are ideally suited to flow cytometric analysis and can be used and detected by flow cytometry to provide information about the physiological status of algal cells in response to toxicants.
Keywords:	aquatic environment; ecotoxicological models; heavy metals; flow cytometry; ecotoxicology
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*Correspondence:	Daniela Bratosin, National Institute for Biological Science Research & Development, Bucharest, Romania; Splaiul Independentei no. 296, district 6, Bucharest, Romania, Tel/Fax +40-(021)-2200881, email:bratosind@yahoo.com