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Development of methods for breeding high-lipidcontent algal strain Chlamydomonas reinhardtii using fluorescence-activated cell sorting
Fedorko, Jan ; Búzová, Diana ; Červený, Jan
Green microalgae are among the most widely distributed microorganisms in the biosphere. They are significant contributors to global photosynthetic productivity and are interesting for biotechnology due to their large variety of high-value compound accumulation and range of applications. To achieve profitable microalgae cultures for biotechnology, one wants to combine antagonistic properties: rapid growth and high accumulation of specific compounds. Here, we focus on development of advanced cultivation strategies and breeding methods applied to the model algae Chlamydomonas reinhardtii for optimized production of lipids. For identification, isolation, and subsequent selection of an optimal subpopulation with high lipid content, we used high-throughput fluorescenceactivated cell sorting in combination with imaging flow cytometry on cells stained with lipid-specific fluorescent dye. We observed that post-sort cell viability was not negatively influenced by external parameters used during the sorting procedure (pressure, light quality and quantity, influence of the sorting electromagnetic field, toxic effects of both fluorescent marker and microfluidic system medium composition).

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