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Consequences of Microbial Interactions with Hydrocarbons, Oils, and Lipids: Production of Fuels and Chemicals pp 87–108Cite as

Screening for Lipids from Marine Microalgae Using Nile Red

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Part of the book series: Handbook of Hydrocarbon and Lipid Microbiology ((HHLM))

Abstract

The fluorescent stain Nile Red has been used extensively for the quantification of lipids in phytoplankton, including microalgae, because it preferentially stains neutral lipids and it is economical and sensitive to use for screening purposes. Although its basic application has not changed for several decades, recent improvements have been made to improve its utility across applications. Here we describe additional refinements in its application and interpretation as a high-throughput method for the rapid quantification of neutral lipids in liquid cultures of marine phytoplankton. Specifically we address (1) interspecies comparisons, (2) fluorescence excitation and emission wavelengths, and (3) the time course of the Nile Red signal in the context of using bulk or cell-specific fluorescence to quantify neutral lipids of live or preserved cells. We show that with proper caution in its interpretation across species and physiological states the quantity of lipid in hundreds of small volume samples can be reliably assessed daily using a refined Nile Red protocol.

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Abbreviations

DAG:

Diacylglycerol

DGCC:

1,2-Diacylglyceryl-3-(O-carboxyhydroxymethylcholine)

DGDG:

Digalactosyl diacylglycerol

DGTA:

1,2-Diacylglyceryl-O-2″-(hydroxymethyl)-(N,N,N-trimethyl)-β-alanine

DGTS:

Diacylgycerol-N-trimethylhomoserine

MGDG:

Monogalactosyl diacylglycerol

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PG:

Phosphatidylgycerol

SQDG:

Sulfoquinovosyl diacylglycerol

TAG:

Triacylglycerol

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Acknowledgments

The authors acknowledge valuable contributions to design of this experiment and critical reading of a prior version of the manuscript by Mark Huntley and Ian Archibald. The authors also thank anonymous reviewers and editors for helpful comments. This work was funded in part by US DOE #DE-EE0007091 and US NSF OCE#14-16665 to ZIJ.

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Authors and Affiliations

  1. Marine Laboratory, Nicholas School of the Environment, and Biology Department, Duke University, 135 Duke Marine Lab Rd, Beaufort, NC, 28516, USA

    Zackary I. Johnson, Sara K. Blinebry, Sarah E. Loftus & Courtney Swink

  2. School of Ocean and Earth Science and Technology, University of Hawaii, 1960 East-West Rd, Honolulu, HI, 96822, USA

    Robert R. Bidigare & Susan L. Brown

  3. Department of Oceanography, Dalhousie University, B3H 4J1, Halifax, NS, Canada

    John J. Cullen

  4. Department of Marine Science, The University of Southern Mississippi, 1020 Balch Boulevard, Stennis Space Center, Kiln, MS, 39529, USA

    Donald G. Redalje

  5. Department of Marine Chemistry and Geochemistry, Woods Hole Oceanographic Institution, Woods Hole, MA, 02543, USA

    Benjamin A. S. Van Mooy

Authors
  1. Zackary I. Johnson
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  3. Sara K. Blinebry
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  4. Susan L. Brown
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  5. John J. Cullen
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  6. Sarah E. Loftus
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  7. Donald G. Redalje
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  8. Courtney Swink
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  9. Benjamin A. S. Van Mooy
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Correspondence to Zackary I. Johnson .

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Editors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea (Republic of)

    Sang Yup Lee

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Johnson, Z.I. et al. (2017). Screening for Lipids from Marine Microalgae Using Nile Red. In: Lee, S. (eds) Consequences of Microbial Interactions with Hydrocarbons, Oils, and Lipids: Production of Fuels and Chemicals. Handbook of Hydrocarbon and Lipid Microbiology . Springer, Cham. https://doi.org/10.1007/978-3-319-50436-0_382

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