Screening for Lipids From Marine Microalgae Using Nile Red

  • Zackary I. Johnson
  • Robert R. Bidigare
  • Sara K. Blinebry
  • Susan L. Brown
  • John J. Cullen
  • Sarah E. Loftus
  • Donald G. Redalje
  • Courtney Swink
  • Benjamin A. S. Van Mooy
Living reference work entry
Part of the Handbook of Hydrocarbon and Lipid Microbiology book series (HHLM)


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.


Neutral Lipid Lipid Class Evaporative Light Scatter Detection Equivalent Carbon Number Microalgae Strain 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.







Digalactosyl diacylglycerol






Monogalactosyl diacylglycerol








Sulfoquinovosyl diacylglycerol





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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Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  • Zackary I. Johnson
    • 1
  • Robert R. Bidigare
    • 2
  • Sara K. Blinebry
    • 1
  • Susan L. Brown
    • 2
  • John J. Cullen
    • 3
  • Sarah E. Loftus
    • 1
  • Donald G. Redalje
    • 4
  • Courtney Swink
    • 1
  • Benjamin A. S. Van Mooy
    • 5
  1. 1.Marine Laboratory, Nicholas School of the Environment, and Biology DepartmentDuke UniversityBeaufortUSA
  2. 2.School of Ocean and Earth Science and TechnologyUniversity of HawaiiHonoluluUSA
  3. 3.Department of OceanographyDalhousie UniversityHalifaxCanada
  4. 4.Department of Marine ScienceThe University of Southern MississippiKilnUSA
  5. 5.Department of Marine Chemistry and GeochemistryWoods Hole Oceanographic InstitutionWoods HoleUSA

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