Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 506642
Title How to us Nile Red, a selective fluorescent stain for microalgal neutral lipids
Author(s) Alemán-Nava, Gibrán S.; Cuellar-Bermudez, Sara P.; Cuaresma, María; Bosma, Rouke; Muylaert, Koenraad; Ritmann, Bruce E.; Parra, Roberto
Source Journal of Microbiological Methods 128 (2016). - ISSN 0167-7012 - p. 74 - 79.
DOI https://doi.org/10.1016/j.mimet.2016.07.011
Department(s) Bioprocess Engineering
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2016
Keyword(s) Biodiesel - Microalgae - Neutral lipids - Nile Red - Protocol
Abstract

The use of Nile Red for rapid monitoring of the neutral lipid content in microalgae has gained interest over the last decade, since neutral lipids are feedstock for renewable transportation fuel. In this review, we discuss the main considerations needed to make an NR protocol reliable for staining neutral lipids in microalgae. Cell wall permeability must be enhanced by using stain carriers: DMSO (5% v/v to 25% v/v), glycerol (0.1 to 0.125 mg/mL), or EDTA (3.0 to 3.8 mg/mL). Temperatures between 30 and 40 °C facilitate the diffusion of NR through the cell wall without incurring excess quenching. Good NR-lipid interaction requires using a low NR/cell ratio; the NR concentration must be between 0.25 μg/mL and 2.0 μg/mL, and the cell concentration > 5 × 104 cells/mL. In order to have the maximum and stable NR fluorescence, it is necessary to scan the excitation/emission wavelengths for up to a 40-min of incubation time. We outline a five-step method to customize the Nile Red protocol to a specific strain: 1) Evaluate the strain's suitability by checking for the presence of neutral lipid, 2) Select of the best excitation/emission wavelength, 3) Optimization of incubation time, stain carrier, dye concentration, and temperature, 4) Prepare single-strain algal cultures with different lipid contents to calibrate NR fluorescence with neutral-lipid content, and 5) Correlate NR fluorescence intensity to neutral lipid content for the same strain. Once the protocol is customized, the NR method allows for rapid and reliable monitoring of neutral lipid content of a microalgae strain.

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