How to use Nile Red, a selective fluorescent stain for microalgal neutral lipids

Alemán‐Nava, Gibrán S.; Cuéllar‐Bermúdez, Sara P.; Cuaresma, María; Bosma, R.; Muylaert, Koenraad; Ritmann, Bruce E.; Parra‐Saldívar, Roberto

doi:10.1016/j.mimet.2016.07.011

Cited by 62 publications

(35 citation statements)

References 46 publications

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“…Based on literature data we may also consider the role of formulating chemicals, in Calypso particularly, the (10%) glycerol content according to the safety data sheet (MSDS Canada). Glycerol is known to facilitate membrane permeability of lipids and fluorescent dyes without affecting their viability (Doan and Obbard, 2011;Natunen et al, 2015;Aleman-Nava et al, 2016). We cannot exclude a non-selective permeability enhancement resulted by the glycerol content in Calypso leading to higher accumulation of both rhodamine B and the active component, thiacloprid.…”

Section: Resultsmentioning

confidence: 92%

Neonicotinoid insecticides are potential substrates of the multixenobiotic resistance (MXR) mechanism in the non-target invertebrate, Dreissena sp.

et al. 2018

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Mussels are among the most frequently used invertebrate animals in aquatic toxicology to detect toxic exposure in the environment. The presence and activity of a cellular defence system, the multixenobiotic resistance (MXR) mechanism, was also established in these organisms.In isolated gill tissues of dreissenid mussels (D. bugensis) the MXR activity was assayed after treatment by commercially available insecticides (formulated products) which contain neonicotinoids as their active ingredients: Actara (thiamethoxam), Apacs (clothianidin), Calypso (thiacloprid) and Kohinor (imidacloprid), respectively. While applying the accumulation assay method, 0.5 μM rhodamine B was used as model substrate and 20 μM verapamil as model inhibitor of the MXR mechanism.In acute (in vitro) experiments when isolated gills were co-incubated in graded concentrations of insecticides and rhodamine B simultaneously, Calypso and Kohinor treatment resulted increasing rhodamine accumulation. Chemical analysis of gills in vitro incubated in insecticides demonstrated higher tissue concentrations of thiamethoxam, clothianidin and thiacloprid in the presence of verapamil suggesting that the active ingredients of Actara, Apacs and Calypso are potential substrates of the MXR mediated cellular efflux. In contrast, verapamil did significantly alter the accumulated imidacloprid concentrations in gills, suggesting that the active component of Kohinor is not transported by the MXR mechanism.Chronic (in vivo) exposures of the intact animals in lower, 1, 10 mg/L concentration of neonicotinoid products, resulted in a decreased level of both rhodamine accumulation and verapamil inhibition by the 12 th -14 th days of treatment. These results suggest an enhancement of MXR activity (chemostimulation), building up gradually in the animals exposed to Actara, Apacs and Kohinor, respectively.Neonicotinoid-type insecticides are generally considered as selective neurotoxins for insects, targeting the nicotinic type acetylcholine receptors (nAChRs) in their central nervous system. Our present results provide the first evidences that neonicotinoid insecticides are also able to alter the transmembrane transport mechanisms related to the MXR system.

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Section: Resultsmentioning

confidence: 92%

Neonicotinoid insecticides are potential substrates of the multixenobiotic resistance (MXR) mechanism in the non-target invertebrate, Dreissena sp.

et al. 2018

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“…To this end, several alternative techniques enabling high-throughput and reliable lipid quantification have emerged recently [31,33]. Fluorescence spectroscopy is considered the simplest and most cost-effective alternative to gravimetric methods for lipid quantification, and fluorescence detection methods have been developed for direct lipid measurements [34][35][36]. Fluorescent dyes such as Nile Red and boron-dipyrromethene (BODIPY) 505/515, which interact with intracellular lipids and emit fluorescence signals correlated to lipid content, are commonly used in such assays [31,37].…”

Section: Sucrose 49mentioning

confidence: 99%

“…Fluorescent dyes such as Nile Red and boron-dipyrromethene (BODIPY) 505/515, which interact with intracellular lipids and emit fluorescence signals correlated to lipid content, are commonly used in such assays [31,37]. These methods, based on a vital dye, offer a rapid and inexpensive approach for measuring lipid content and can be applied in various systems, in situ and in vitro [35]. Other non-destructive methods, such as infrared (IR), nuclear magnetic resonance (NMR), and Raman spectroscopy, have also been used to some extent for lipid determination in oleaginous microorganisms [38][39][40][41][42].…”

Section: Sucrose 49mentioning

confidence: 99%

“…Other non-destructive methods, such as infrared (IR), nuclear magnetic resonance (NMR), and Raman spectroscopy, have also been used to some extent for lipid determination in oleaginous microorganisms [38][39][40][41][42]. Techniques for lipid content determination in microalgae have been thoroughly described in several recent reviews [31,[33][34][35] and are also commonly applied to lipid quantification in other oleaginous microorganisms. Other important factors, such as the degree of unsaturation and saturation, the position and number of double bonds in the hydrocarbon chain, the amount of free fatty acids, TAGs, diacylglycerols (DAGs), monoacylglycerols (MAGs), and steryl esters, should also be taken into consideration when evaluating microbial lipids for biofuels [43].…”

Section: Sucrose 49mentioning

confidence: 99%

“…The interaction between Nile Red and lipids can be further improved by using a low dye:cell ratio (0.25-2.0 μg dye/mL) and cell concentrations > 5 × 10 4 cells/mL. Finally, to determine the maximum Nile Red fluorescence intensity, it is recommended to scan excitation and emission for up to 40 min [35].…”

Section: Fluorescence Spectroscopymentioning

confidence: 99%

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Lipids detection and quantification in oleaginous microorganisms: an overview of the current state of the art

2019

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Oleaginous microorganisms are among the most promising feedstocks for the production of lipids for biofuels and oleochemicals. Lipids are synthesized in intracellular compartments in the form of lipid droplets. Therefore, their qualitative and quantitative analysis requires an initial pretreatment step that allows their extraction. Lipid extraction techniques vary with the type of microorganism but, in general, the presence of an outer membrane or cell wall limits their recovery. This review discusses the various types of oleaginous microorganisms, their lipid accumulating capabilities, lipid extraction techniques, and the pretreatment of cellular biomass for enhanced lipid recovery. Conventional methods for lipid quantification include gravimetric and chromatographic approaches; whereas nonconventional methods are based on infrared, Raman, nuclear magnetic resonance, and fluorescence spectroscopic analysis. Recent advances in these methods, their limitations, and fields of application are discussed, with the aim of providing a guide for selecting the best method or combination of methods for lipid quantification.

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Temperature and nutrient coupled stress on microalgal neutral lipids for low‐carbon fuels production

Anandraj

White

Bwapwa

2021

Biofuels Bioprod Bioref

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A freshwater Chlorella sp. was cultured at temperatures from 10 to 45 °C and synergistic stress was employed by using selective and complete nutrient depleted growth media. Temperature fluctuations were found to have a significant effect on the microalgae physiology. The optimum temperature of 35 °C shows suitability for growth in a sub‐tropical climate. The fatty acid methyl esters’ (FAMEs') composition was significantly altered when cultures were incubated under various pre‐determined stress conditions. The effect of dual stress was found to have the greatest increase in the lipid yield, averaging 55% (w/w). The majority of the identified FAMEs (C16 and C18) showed the potential for quality biodiesel and aviation fuel due to the characteristics of FAME. This study is relevant for commercial‐scale biodiesel production and the technologies developed would improve the economic feasibility and potential of microalgal biomass feedstock for biodiesel production by transesterification. There is also high potential for low‐carbon jet fuel by thermal cracking or pyrolysis and fractionation of crude‐bio‐oil from Chlorella sp. These processes will convert carbon chains or FAMES in the C14 to C16 range, which is appropriate for low‐carbon jet fuel. It is therefore possible to generate biodiesel and aviation fuel from Chlorella sp. cultivated in freshwater at achievable temperatures in a sub‐tropical environment under total nutrient deprivation, aiming to stimulate a bio‐oil increase. © 2021 Society of Industrial Chemistry and John Wiley & Sons Ltd.

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How to use Nile Red, a selective fluorescent stain for microalgal neutral lipids

Cited by 62 publications

References 46 publications

Neonicotinoid insecticides are potential substrates of the multixenobiotic resistance (MXR) mechanism in the non-target invertebrate, Dreissena sp.

Neonicotinoid insecticides are potential substrates of the multixenobiotic resistance (MXR) mechanism in the non-target invertebrate, Dreissena sp.

Lipids detection and quantification in oleaginous microorganisms: an overview of the current state of the art

Temperature and nutrient coupled stress on microalgal neutral lipids for low‐carbon fuels production

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