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

Katariina Natunen, Jukka Seppälä, Riikka-Juulia Koivula, Jukka Pellinen

Journal of phycology 2017 Apr

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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. © 2016 Phycological Society of America.

Citation

Katariina Natunen, Jukka Seppälä, Riikka-Juulia Koivula, Jukka Pellinen. Monitoring cell-specific neutral lipid accumulation in Phaeodactylum tricornutum (Bacillariophyceae) with Nile Red staining - a new method for FlowCAM. Journal of phycology. 2017 Apr;53(2):396-404