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Single Cell Analysis of Microalgae and Associated Bacteria Flora by Using Flow Cytometry

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Abstract

Microalgae are usually cultivated in mixed cultures with associated bacteria flora. Despite bacteria contaminants can strongly influence microalgal growth, they are rarely monitored, mainly because standardized and quick methods are missing. The aim of this study was to develop a quick flow cytometric method to quantify coenobia-forming microalgae (Tetradesmus obliquus) and associated bacteria. Staining conditions and sonication pretreatment were optimized to obtain the most accurate microalgal and bacterial cell counting. To pre-treat microalgae before counting, pulsed sonication (ton/toff = 30/60) was better than continuous sonication since it allows obtaining 100% single cells by minimizing cell lysis. On the contrary, sonication was not required for bacteria, because they were mainly found as single cells and only a relevant cell lysis was obtained when sonication was applied. To analyze samples as those tested in this study, bacteria should be analyzed directly after fixation and DNA staining, without sonication (duration of analysis: 90 min). Instead, sonication was mandatory for microalgae, while fixation and DNA staining could be avoided (duration of analysis: 30 min). Future studies should investigate the effect of the biological variability, that seems to be the most relevant factor affecting the accuracy and reproducibility of the flow cytometric analysis.

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Change history

  • 22 January 2021

    In the original upload, the revised date is written as 11 April 2011, It should be 2021.

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This research did not receive external funding.

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Authors and Affiliations

  1. Department of Chemistry, University Sapienza of Rome, Piazzale Aldo Moro 5, Rome, 00185, Italy

    Fabrizio Di Caprio, Simone Posani, Pietro Altimari & Francesca Pagnanelli

  2. Department of Mechanical, Chemical and Materials Engineering, University of Cagliari, Via Marengo 2, Cagliari, 09123, Italy

    Alessandro Concas

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  1. Fabrizio Di Caprio
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  2. Simone Posani
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  3. Pietro Altimari
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  4. Alessandro Concas
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  5. Francesca Pagnanelli
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Correspondence to Fabrizio Di Caprio.

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Di Caprio, F., Posani, S., Altimari, P. et al. Single Cell Analysis of Microalgae and Associated Bacteria Flora by Using Flow Cytometry. Biotechnol Bioproc E 26, 898–909 (2021). https://doi.org/10.1007/s12257-021-0054-9

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  • DOI: https://doi.org/10.1007/s12257-021-0054-9

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