Nile Red staining of phytoplankton neutral lipids: species-specific fluorescence kinetics in various solvents

Natunen, Katariina; Seppälä, Jukka; Schwenk, Dagmar; Rischer, Heiko; Spilling, Kristian; Tamminen, Timo

doi:10.1007/s10811-014-0404-5

Cited by 29 publications

(23 citation statements)

References 22 publications

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“…These methods are more efficient, cheaper and easier to use compared to all of the previously mentioned techniques. Additionally, they are significantly time saving (Govender et al 2012;Ren et al 2013;Mishra et al 2014;Cabanelas et al 2015;Natunen et al 2015).…”

Section: Introductionmentioning

confidence: 99%

Imaging the accumulated intracellular microalgal lipids as a response to temperature stress

et al. 2017

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Over the last few decades, many scientists considered microalgae as promising actors for future biofuels because of the high lipid productivity inside their cells. Moreover, much attention has been paid to algal lipids as they can be used in biodiesel production. In this study, we optimized the different suitable conditions such as incubation time, incubation temperature, Dimethylesulfoxide and Nile red concentrations of the lipophilic fluorescence dye Nile red as an excellent and fast vital stain to detect and quantify intracellular lipids. This was achieved using the green alga Nannochloropsis salina. In addition, investigating the accumulation of lipid vesicles inside different isolated microalgal species as a response to temperature stress. Furthermore, the confocal laser scanning microscopy (LS510) for imaging and measuring the size and volume of the accumulated lipid vesicles was used.

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Section: Introductionmentioning

confidence: 99%

Imaging the accumulated intracellular microalgal lipids as a response to temperature stress

et al. 2017

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“…, Natunen et al. ). Phaeodactylum tricornutum was cultivated in two parallel polycarbonate bottles (Cultures 1 and 2) using two liters of modified f/2 medium (Guillard ; salinity 6 g · L −1 ) that had been adjusted to the molar ratio N:P = 4 (i.e., [N] = 290, [P] = 73, [Si] = 145 μmol · L −1 ), so that the cultures would become nitrogen‐limited during growth and start accumulating neutral lipids.…”

Section: Methodsmentioning

confidence: 98%

“…Based on previous findings (Natunen et al. ), to duplicate 2.4 mL culture samples from each cultivation bottle (Culture 1 and 2), 0.6 mL of NR (for microscopy, Sigma‐Aldrich Corp., St. Louis, MO, USA; stock solution 0.25 mg · mL −1 ) in DMSO (Sigma‐Aldrich Corp., St. Louis, MO, USA) was added, so that a final NR concentration of 1 μg · mL −1 , and a DMSO concentration of 20% (v/v) in the analyzed sample were obtained. The samples were then incubated for 13 min (Natunen et al.…”

Section: Methodsmentioning

confidence: 99%

“…, Natunen et al. ). In case of poor cell penetration of the dye, organic solvents, such as DMSO, have proven to be helpful with some species (Chen et al.…”

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confidence: 98%

“…, Natunen et al. ). It also needs to be noted that the NR fluorescence measured from a stained sample is always relative and system‐specific, so for absolute quantification of lipids with the NR method, a calibration step is compulsory.…”

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confidence: 98%

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Monitoring cell‐specific neutral lipid accumulation in Phaeodactylum tricornutum (Bacillariophyceae) with Nile Red staining – a new method for FlowCAM

Natunen

Seppälä

Koivula

et al. 2017

Journal of Phycology

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With the fluorescent stain Nile Red (NR), phytoplankton lipid accumulation can be monitored quickly and in situ. In the light of recent results in phytoplankton diversity research, there is also a need for cell- and species-specific lipid measurement techniques. The objective of this work was to investigate whether cell-specific phytoplankton lipid accumulation could be monitored with the image-based particle analyzer FlowCAM™ and NR staining. Applying Phaeodactylum tricornutum as a model species, we compared the FlowCAM method to two established lipid quantification methods: spectrofluorometric NR fluorescence measurement and total lipid analysis by gas chromatography. The experiment was carried out in batch cultures under nitrogen limitation to induce lipid accumulation. We showed significant correlation between the three different lipid quantification methods confirming the applicability of the novel FlowCAM method in cell-specific and near real-time lipid quantification. Furthermore, with the method described here, the lipid content of taxonomically distinguished cells can eventually be measured from multispecies cultures, opening several new possibilities to study species-specific responses to stress conditions and the complementarity effect.

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Lipid quantification techniques for screening oleaginous species of microalgae for biofuel production

Hounslow

Noirel

Gilmour

et al. 2016

Euro J Lipid Sci & Tech

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This review examines the available literature on quantifying lipids in microalgae suitable for biofuels research. It discusses their advantages and disadvantages, their prevalence in the literature, and draws conclusions about the best way to approach choosing a suitable lipid measurement technique for microalgal biofuels research. We conclude that the method must be chosen based on the following key criteria: (1) the level of detail required in the results, and (2) the amount of biomass that can be spared for the assay. This review establishes that no method can be used as a "golden standard" for all microalgae. However, we present a systematic decision chart to choose the best measurement method or combination of methods to provide a guide to those wishing to understand the differences between the myriad of lipid measurement techniques.Practical applications: This review will allow researchers new to the field to choose the most appropriate techniques for quantifying lipids in the microalgal species under study. The review is also intended to act as a gateway to the wider literature and will enable researchers to look in depth at a particular technique before carrying out experimental work.

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Nile Red staining of phytoplankton neutral lipids: species-specific fluorescence kinetics in various solvents

Cited by 29 publications

References 22 publications

Imaging the accumulated intracellular microalgal lipids as a response to temperature stress

Imaging the accumulated intracellular microalgal lipids as a response to temperature stress

Monitoring cell‐specific neutral lipid accumulation in Phaeodactylum tricornutum (Bacillariophyceae) with Nile Red staining – a new method for FlowCAM

Lipid quantification techniques for screening oleaginous species of microalgae for biofuel production

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