title: A simple and rapid protocol for measuring neutral lipids in algal cells using fluorescence.
abstract: Algae are considered excellent candidates for renewable fuel sources due to their natural lipid storage capabilities. Robust monitoring of algal fermentation processes and screening for new oil-rich strains requires a fast and reliable protocol for determination of intracellular lipid content. Current practices rely largely on gravimetric methods to determine oil content, techniques developed decades ago that are time consuming and require large sample volumes. In this paper, Nile Red, a fluorescent dye that has been used to identify the presence of lipid bodies in numerous types of organisms, is incorporated into a simple, fast, and reliable protocol for measuring the neutral lipid content of Auxenochlorella protothecoides, a green alga. The method uses ethanol, a relatively mild solvent, to permeabilize the cell membrane before staining and a 96 well micro-plate to increase sample capacity during fluorescence intensity measurements. It has been designed with the specific application of monitoring bioprocess performance. Previously dried samples or live samples from a growing culture can be used in the assay.
authors: Department of Chemical and Materials Engineering, University of Alberta.
authors: Zachary J
authors: ZJ
authors: Storms
authors: Department of Chemical and Materials Engineering, University of Alberta.
authors: Elliot
authors: E
authors: Cameron
authors: Department of Chemical and Materials Engineering, University of Alberta; Department of Chemical and Petroleum Engineering, University of Calgary.
authors: Hector
authors: H
authors: de la Hoz Siegler
authors: Department of Chemical and Materials Engineering, University of Alberta; wcm@ualberta.ca.
authors: William C
authors: WC
authors: McCaffrey
mesh_terms: D018533:Biomass; D048408:Chlorella vulgaris / Q000737:chemistry*; D005470:Fluorometry / Q000379:methods; D008055:Lipids / Q000032:analysis*; D059625:Liquid-Liquid Extraction / Q000379:methods; D058086:Microalgae / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods; D010078:Oxazines / Q000737:chemistry; D044444:Scenedesmus / Q000737:chemistry*
chemical_list: D008055:Lipids; D010078:Oxazines; C044808:nile red