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Flow cytometry sorting of freshwater phytoplankton

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Abstract

We used the flow sorting capacities of a benchtop FACSCalibur flow cytometer to analyze the phytoplankton community of four different aquatic ecosystems. We show that despite the high optical, mechanistic, and hydrodynamic stress for the cells while sorted, most of the targeted populations could be isolated and grew in mixed culture media subsequent to sorting. Forty-five phytoplankton taxa were isolated, including green algae (29 species), cyanobacteria (eight), diatoms (seven), and cryptomonads (one). The isolation success average was high since 80% of the total sorted populations grew successfully and 47% constituted monocultures. It is noteworthy, however, that some groups could not be isolated, as for example colonial cyanobacteria, chrysophytes, euglenophytes, desmids, or dinoflagellates, and some species such as Cryptomonas sp. were very sensitive to the sorting process. It is proposed that flow cytometric analysis of freshwater phytoplankton might be a relevant tool for water managers and could be applied in some specific cases, such as early monitoring of blooming taxa or basic bio-monitorings of key species. The higher isolation average obtained from the flow sorting can also be powerful for the physiological or molecular study of some taxa after their cultivation.

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Acknowledgments

We are grateful to Pascal Perney, Jean-Christophe Hustache, Pascal Chifflet (INRA Thonon), and Gérard Paolini (CISALB, Aix-les-Bains), who realized the sampling of the three alpine lakes during the period of this study. Gérard Beaudoin and Yannik Guillemin (IIBRBS) are acknowledged for their help in the sampling of the Reservoir Marne. We want to thank Eliane Menthon for the preparation of the culture media and Jean Claude Druart (INRA, Thonon) for his support in the taxonomic phytoplankton identification.

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  1. Station d’Hydrobiologie Lacustre, INRA, UMR CARRTEL, 75 Avenue de Corzent, 74203, Thonon-les-Bains Cedex, France

    Maria Cellamare, Anne Rolland & Stéphan Jacquet

  2. Cemagref, UR REBX, 50 avenue de Verdun, 33612, Cestas Cedex, France

    Maria Cellamare

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  1. Maria Cellamare
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Cellamare, M., Rolland, A. & Jacquet, S. Flow cytometry sorting of freshwater phytoplankton. J Appl Phycol 22, 87–100 (2010). https://doi.org/10.1007/s10811-009-9439-4

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