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BODIPY vital staining as a tool for flow cytometric monitoring of intracellular lipid accumulation in Nannochloropsis gaditana

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Abstract

The monitoring of lipid accumulation in microalgae is a key issue that needs an effective and fast method of analysis, mainly when biofuel-directed cultures are carried out. Boron-dipyrromethene (BODIPY), a green fluorescent lipophilic dye, was employed combined with flow cytometry for monitoring lipid storage in Nannochloropsis gaditana, an oleaginous marine microalga. Two growth conditions, nitrogen sufficiency and nitrogen starvation, were tested for evaluation of lipid accumulation. The removal of nitrogen from growth media resulted in a reduction in the chloroplast volume and the appearance of large intracellular lipid bodies compared with cells grown in nitrogen-sufficient media. Three concentrations of BODIPY were tested for optimization of the BODIPY lipid staining in microalgae: 1, 50 and 250 ng mL−1. A concentration of 1 ng mL−1 for percentage of fluorescent cell analysis and 50 ng mL−1 for mean fluorescence intensity analysis were shown to be optimal. BODIPY staining data showed results similar to the results obtained from the conventional gravimetric method. Application of the BODIPY staining together with flow cytometric analysis is a simple, rapid and reliable method for in situ monitoring of lipid accumulation in N. gaditana.

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Acknowledgments

This study was conducted with the support of the Basque Government, through the EMAITEK program.

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Authors and Affiliations

  1. GAIKER Technology Centre, Parque Tecnológico Ed 202, Zamudio, 48170, Vizcaya, Spain

    Vanessa Benito, Felipe Goñi-de-Cerio & Pilar Brettes

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  1. Vanessa Benito
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  2. Felipe Goñi-de-Cerio
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  3. Pilar Brettes
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Correspondence to Vanessa Benito.

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Benito, V., Goñi-de-Cerio, F. & Brettes, P. BODIPY vital staining as a tool for flow cytometric monitoring of intracellular lipid accumulation in Nannochloropsis gaditana . J Appl Phycol 27, 233–241 (2015). https://doi.org/10.1007/s10811-014-0310-x

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  • DOI: https://doi.org/10.1007/s10811-014-0310-x

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