Visualizing "green oil" in live algal cells

Cooper, Mark S.; Hardin, William; Petersen, Timothy W.; Cattolico, Rose Ann

doi:10.1016/j.jbiosc.2009.08.004

Cited by 195 publications

(140 citation statements)

References 16 publications

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“…BODIPY 505/515 (Invitrogen, Carlsbad, CA, USA) serving as an excellent stain for lipid bodies of live algal cells was used in this study. BODIPY 505/515 staining was conducted according to the protocol described by Cooper et al (2010). The cells were observed and photographed under the Olympus BX41 microscope.…”

Section: Lipid Extraction and Analysismentioning

confidence: 99%

Scenedesmus sp. NJ-1 isolated from Antarctica: a suitable renewable lipid source for biodiesel production

Chen

Gong

Fang

et al. 2012

World J Microbiol Biotechnol

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Microalgal lipids are promising alternative feedstocks for biodiesel production. Scenedesmus sp. NJ-1, an oil-rich freshwater microalga isolated from Antarctica, was identified to be a suitable candidate to produce biodiesel in this study. This strain could grow at temperatures ranging from 4 to 35°C. With regular decrease in nitrate concentration in the medium, large quantities of triacylglycerols accumulated under batch culture conditions detected by thin layer chromatography and BODIPY 505/515 fluorescent staining. Scenedesmus sp. NJ-1 achieved the average biomass productivity of 0.105 g l -1 d -1 (dry weight) and nearly the highest lipid content (35 % of dry cell weight) was reached at day 28 in the batch culture. Neutral lipids accounted for 78 % of total lipids, and C18:1 (n-9), C16:0 were the major fatty acids in total lipids, composing 37 and 20 % of total fatty acids of Scenedesmus sp. NJ-1 grown for 36 days, respectively. These results suggested that Scenedesmus sp. NJ-1 was a good source of microalgal oils for biodiesel production.

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Section: Lipid Extraction and Analysismentioning

confidence: 99%

Scenedesmus sp. NJ-1 isolated from Antarctica: a suitable renewable lipid source for biodiesel production

Chen

Gong

Fang

et al. 2012

World J Microbiol Biotechnol

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“…Fluorescence microscopy is the most widespread technique, relying on lipid-specific fluorescent markers: Nile Red is commonly used, but its poor permeability through the cell walls causes difficulties, as does its nonspecific binding (Chen et al, 2009). Other fluorophores like BODIPY 505/515 also have been suggested for live-cell studies (Cooper et al, 2010;Wong and Franz, 2013). Still, invasive techniques are needed, requiring solvents to facilitate the transport of the labeling molecules into the cell, potentially inducing stress responses that may affect cell metabolism.…”

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confidence: 99%

Imaging of Lipids in Microalgae with Coherent Anti-Stokes Raman Scattering Microscopy

et al. 2015

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Microalgae have great prospects as a sustainable resource of lipids for refinement into nutraceuticals and biodiesel, which increases the need for detailed insights into their intracellular lipid synthesis/storage mechanisms. As an alternative strategy to solvent-and label-based lipid quantification techniques, we introduce time-gated coherent anti-Stokes Raman scattering (CARS) microscopy for monitoring lipid contents in living algae, despite strong autofluorescence from the chloroplasts, at approximately picogram and subcellular levels by probing inherent molecular vibrations. Intracellular lipid droplet synthesis was followed in Phaeodactylum tricornutum algae grown under (1) light/nutrient-replete (control [Ctrl]), (2) light-limited (LL), and (3) nitrogenstarved (NS) conditions. Good correlation (r 2 = 0.924) was found between lipid volume data yielded by CARS microscopy and total fatty acid content obtained from gas chromatography-mass spectrometry analysis. In Ctrl and LL cells, micron-sized lipid droplets were found to increase in number throughout the growth phases, particularly in the stationary phase. During more excessive lipid accumulation, as observed in NS cells, promising commercial harvest as biofuels and nutritional lipids, several micron-sized droplets were present already initially during cultivation, which then fused into a single giant droplet toward stationary phase alongside with new droplets emerging. CARS microspectroscopy further indicated lower lipid fluidity in NS cells than in Ctrl and LL cells, potentially due to higher fatty acid saturation. This agreed with the fatty acid profiles gathered by gas chromatography-mass spectrometry. CARS microscopy could thus provide quantitative and semiqualitative data at the singlecell level along with important insights into lipid-accumulating mechanisms, here revealing two different modes for normal and excessive lipid accumulation.

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“…Other commonly used methods to determine the lipid content in microalgae include staining the biomass with lipophilic fluorescent stains such as Nile red or BODIPY and measuring the fluorescence signal 21,22 . An advantage of these methods is that they are less laborious than alternative methods.…”

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confidence: 99%

Analysis of Fatty Acid Content and Composition in Microalgae

Breuer

Evers

Vree

et al. 2013

JoVE

118

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A method to determine the content and composition of total fatty acids present in microalgae is described. Fatty acids are a major constituent of microalgal biomass. These fatty acids can be present in different acyl-lipid classes. Especially the fatty acids present in triacylglycerol (TAG) are of commercial interest, because they can be used for production of transportation fuels, bulk chemicals, nutraceuticals (ω-3 fatty acids), and food commodities. To develop commercial applications, reliable analytical methods for quantification of fatty acid content and composition are needed. Microalgae are single cells surrounded by a rigid cell wall. A fatty acid analysis method should provide sufficient cell disruption to liberate all acyl lipids and the extraction procedure used should be able to extract all acyl lipid classes.With the method presented here all fatty acids present in microalgae can be accurately and reproducibly identified and quantified using small amounts of sample (5 mg) independent of their chain length, degree of unsaturation, or the lipid class they are part of. This method does not provide information about the relative abundance of different lipid classes, but can be extended to separate lipid classes from each other.The method is based on a sequence of mechanical cell disruption, solvent based lipid extraction, transesterification of fatty acids to fatty acid methyl esters (FAMEs), and quantification and identification of FAMEs using gas chromatography (GC-FID). A TAG internal standard (tripentadecanoin) is added prior to the analytical procedure to correct for losses during extraction and incomplete transesterification. Video LinkThe video component of this article can be found at

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Visualizing "green oil" in live algal cells

Cited by 195 publications

References 16 publications

Scenedesmus sp. NJ-1 isolated from Antarctica: a suitable renewable lipid source for biodiesel production

Scenedesmus sp. NJ-1 isolated from Antarctica: a suitable renewable lipid source for biodiesel production

Imaging of Lipids in Microalgae with Coherent Anti-Stokes Raman Scattering Microscopy

Analysis of Fatty Acid Content and Composition in Microalgae

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