Imaging of Lipids in Microalgae with Coherent Anti-Stokes Raman Scattering Microscopy1[OPEN]

@article{Cavonius2015ImagingOL,
  title={Imaging of Lipids in Microalgae with Coherent Anti-Stokes Raman Scattering Microscopy1[OPEN]},
  author={Lillie R. Cavonius and Helen Fink and Jūris Ki{\vs}kis and Eva Albers and Ingrid Undeland and Annika M K Enejder},
  journal={Plant Physiology},
  year={2015},
  volume={167},
  pages={603 - 616}
}
Lipids are accumulated as giant droplets alongside coalescing emerging droplets under excessive lipid storage, in contrast to the multiple micron-sized droplets formed at normal conditions. 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… 

Figures and Tables from this paper

Label-free in vivo analysis of intracellular lipid droplets in the oleaginous microalga Monoraphidium neglectum by coherent Raman scattering microscopy
TLDR
The lipid profile during cellular adaption to nitrogen starvation showed a two-phase characteristic with initially fatty acid recycling and subsequent de novo lipid synthesis, which demonstrates the potential of quantitative CARS microscopy as a label-free lipid analysis technique for any microalgal species.
Investigation of Microalgal Carotenoid Content Using Coherent Anti-Stokes Raman Scattering (CARS) Microscopy and Spontaneous Raman Spectroscopy.
TLDR
CARS microscopy and Raman microspectroscopy present themselves as viable alternatives to the standard techniques currently in use because of their non-disruptive nature and the wealth of complementary information that could be obtained from them in regards to assessing the chemical content of microalgae.
High-speed stimulated Raman scattering microscopy for studying the metabolic diversity of motile Euglena gracilis
TLDR
The SRS system’s fast imaging capability enables quantification and analysis of previously unresolvable cell-to-cell variations in the metabolite accumulation of large motile E. gracilis cell populations.
Non-invasive lipid quantification of living microalgal cultures with digital holographic microscopy
TLDR
A fast quantitative method to assess the evolution of the lipid content inside the diatom Phaeodactylum tricornutum living cells using a specific processing of recorded hologram sequences based on the refocusing capability of digital holographic microscopy.
Revealing Lipid Body Formation and its Subcellular Reorganization in Oleaginous Microalgae Using Correlative Optical Microscopy and Infrared Nanospectroscopy
TLDR
An integrated imaging approach to characterize without labeling at the sub-cellular level the formation of lipid body droplets in microalgae undergoing nitrogen starvation in Parachlorella kesslerii using an innovative infrared nanospectroscopy technique.
Label-free non-invasive quantitative measurement of lipid contents in individual microalgal cells using refractive index tomography
TLDR
This work demonstrates label-free, non-invasive, rapid quantification of the lipid contents in individual micro-algal cells measuring the three-dimensional refractive index tomograms and finds that lipid droplets are identifiable in tomograms by their highRefractive index.
Detection and imaging of lipids of Scenedesmus obliquus based on confocal Raman microspectroscopy
TLDR
There was significant correlation between the fatty acid content measured by the gravimetric method and the integral Raman intensity of the oil characteristic peak measured by Raman spectroscopy, which means that Raman Spectroscopy is applicable to semi-quantitative detection of microalgal lipid content.
...
...

References

SHOWING 1-10 OF 44 REFERENCES
Coherent anti-Stokes Raman scattering and spontaneous Raman spectroscopy and microscopy of microalgae with nitrogen depletion
TLDR
Both spontaneous Raman spectroscopy and coherent anti-Stokes Raman scattering microscopy were found to be suitable analysis tools for microalgae.
Coherent Anti-Stokes Raman Scattering Microscopy of Cellular Lipid Storage
TLDR
Coherent anti-Stokes Raman scattering has a strong potential to become a central instrument for in vivo studies of the lipid metabolism at cellular level, improving present understanding of the mechanisms behind the many metabolism-related diseases, the impact of natural bioactive components in foods, and supporting the development of efficient pharmaceuticals as well as bioengineering processes exploiting the metabolism of microorganisms for the production of alternative energy sources.
Quantitative label-free imaging of lipid composition and packing of individual cellular lipid droplets using multiplex CARS microscopy.
TLDR
A label-free quantitative method is used to image lipids within LDs in 3T3-L1 cells and reveals variations in lipid composition and physical state between LDs contained in the same cell, and even within a single LD.
Microfluidic cytometer for high-throughput measurement of photosynthetic characteristics and lipid accumulation in individual algal cells.
TLDR
A microfluidic cytometer is developed that is believed to be the first time that both photosynthetic efficiency and lipid accumulation have been simultaneously evaluated at the single cell level, and in doing so, the diversity within these populations was revealed.
RNA Interference Silencing of a Major Lipid Droplet Protein Affects Lipid Droplet Size in Chlamydomonas reinhardtii
TLDR
Focusing on the model green alga Chlamydomonas reinhardtii, the accumulation of triacylglycerols and the formation of lipid droplets during nitrogen deprivation were investigated and a major protein was tentatively designated major lipid droplet protein (MLDP), specific to the green algal lineage of photosynthetic organisms.
A comparison of lipid storage in Phaeodactylum tricornutum and Tetraselmis suecica using laser scanning confocal microscopy.
Characterization of major lipid droplet proteins from Dunaliella
TLDR
The isolation of MLDP from three species of the extreme halotolerant green algae Dunaliella identified a conserved 4-proline domain that may be considered as a signature domain of Volvocales green algae MLDP.
Selective imaging of saturated and unsaturated lipids by wide-field CARS-microscopy.
TLDR
The high signal-to-noise ratio of wide-field CARS microscopy that is achieved by an excitation geometry involving a "sheet-of-light"-type illumination is drawn for Quantitative CARS imaging at the =C-H resonance.
A Lipid Droplet Protein of Nannochloropsis with Functions Partially Analogous to Plant Oleosins1[W][OA]
TLDR
Although LDSP presence in Arabidopsis predictably affected LD size, it could not reverse the physiological impact of OLEOSIN deficiency on triacylglycerol hydrolysis during germination and was found to have unique primary sequence but structural similarities to other LD proteins.
...
...