Imaging of Lipids in Microalgae with Coherent Anti-Stokes Raman Scattering Microscopy
Open accessAbstract
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) nitrogen-starved (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 single-cell level along with important insights into lipid-accumulating mechanisms, here revealing two different modes for normal and excessive lipid accumulation.
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The lipid droplet (LD) size of Monoraphidium neglectum is far smaller than the LDs of ≥8 μm3 reported for Phaeodactylum tricornutum.
“which is far smaller than the LDs of ≥8 μm3 reported for the well characterized diatom species P. tricornutum25.”
From Label-free in vivo analysis of intracellular lipid droplets in the oleaginous microalga Monoraphidium neglectum by coherent Raman scattering microscopy (2016) · cites this paper as [25]
CARS microscopy was applied to the diatom Phaeodactylum tricornutum, and the CARS signal was separated from two-photon excited chlorophyll fluorescence by time-gated detection; a correlation between quantitative CARS data and biochemically acquired data was observed.
“Subsequently, CARS microscopy was applied for the diatom Phaeodactylum tricornutum, and the CARS signal was separated from the two-photon excited chlorophyll fluorescence by time-gated detection25. The authors formulated a model relating their quantitative CARS data with biochemically acquired data, and a correlation between these two was observed25.”
From Label-free in vivo analysis of intracellular lipid droplets in the oleaginous microalga Monoraphidium neglectum by coherent Raman scattering microscopy (2016) · cites this paper as [25]
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Cites
Chisti, Y. (2007) · Biotechnology Advances · doi:10.1016/j.biotechadv.2007.02.001 · Cited by 9,524 · see citing works
Andersen, R. A., & Phycological Society of America (2005) · Choice Reviews Online · doi:10.5860/choice.43-0925 · Cited by 1,299 · see citing works
Coherent Anti-Stokes Raman Scattering Microscopy: Instrumentation, Theory, and Applications
Cheng, J., & Xie, X. S. (2003) · The Journal of Physical Chemistry B · doi:10.1021/jp035693v · Cited by 962 · see citing works
A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae
Chen, W., Zhang, C., Song, L., Sommerfeld, M., & Hu, Q. (2009) · Journal of Microbiological Methods · doi:10.1016/j.mimet.2009.01.001 · Cited by 681 · see citing works
Bolton, J. J. (2006) · Journal of Experimental Marine Biology and Ecology · doi:10.1016/j.jembe.2006.05.002 · Cited by 644 · see citing works
Breuer, G., Lamers, P. P., Martens, D. E., Draaisma, R. B., & Wijffels, R. H. (2012) · Bioresource Technology · doi:10.1016/j.biortech.2012.08.003 · Cited by 626 · see citing works
The Lipid-Droplet Proteome Reveals that Droplets Are a Protein-Storage Depot
Cermelli, S., Guo, Y., Gross, S. P., & Welte, M. A. (2006) · Current Biology · doi:10.1016/j.cub.2006.07.062 · Cited by 483 · see citing works
Walther, T. C., & Farese, R. V. (2008) · Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids · doi:10.1016/j.bbalip.2008.10.009 · Cited by 478 · see citing works
Health applications of bioactive compounds from marine microalgae
Raposo, M. F. D. J., Morais, R. M. S. C., & Morais, A. M. M. B. (2013) · Life Sciences · doi:10.1016/j.lfs.2013.08.002 · Cited by 412 · see citing works
Moellering, E. R., & Christoph Benning, A. (2009) · Eukaryotic Cell · doi:10.1128/ec.00203-09 · Cited by 401 · see citing works
Photophysics of the carotenoids associated with the xanthophyll cycle in photosynthesis
Frank, H. A., Cua, A., Chynwat, V., Young, A. J., Gosztola, D. J., & Wasielewski, M. R. (1994) · Photosynthesis Research · doi:10.1007/bf02183041 · Cited by 393 · see citing works
The dynamic roles of intracellular lipid droplets: from archaea to mammals
Murphy, D. J. (2011) · PROTOPLASMA · doi:10.1007/s00709-011-0329-7 · Cited by 392 · see citing works
Yongmanitchai, W., & Ward, O. P. (1991) · Applied and Environmental Microbiology · doi:10.1128/aem.57.2.419-425.1991 · Cited by 371 · see citing works
Fan, J., Andre, C., & Xu, C. (2011) · FEBS Letters · doi:10.1016/j.febslet.2011.05.018 · Cited by 347 · see citing works
Griffiths, M. J., van Hille, R. P., & Harrison, S. T. L. (2012) · Journal of Applied Phycology · doi:10.1007/s10811-011-9723-y · Cited by 341 · see citing works
Athenstaedt, K., Zweytick, D., Jandrositz, A., Kohlwein, S. D., & Daum, G. (1999) · Journal of Bacteriology · doi:10.1128/jb.181.20.6441-6448.1999 · Cited by 316 · see citing works
Tonon, T., Harvey, D., Larson, T. R., & Graham, I. A. (2002) · Phytochemistry · doi:10.1016/s0031-9422(02)00201-7 · Cited by 294 · see citing works
Fernández‐Reiriz, M. J., Pérez-Camacho, A., Ferreiro, M. J., Blanco, J., Planas, M., Campos, M. J., & Labarta, U. (1989) · Aquaculture · doi:10.1016/0044-8486(89)90057-4 · Cited by 265 · see citing works
Nitrogen Starvation Induced Oxidative Stress in an Oil-Producing Green Alga Chlorella sorokiniana C3
Zhang, Y., Chen, H., He, C., & Wang, Q. (2013) · PLoS ONE · doi:10.1371/journal.pone.0069225 · Cited by 247 · see citing works
Rinia, H. A., Burger, K. N. J., Bonn, M., & Mueller, M. (2008) · Biophysical Journal · doi:10.1529/biophysj.108.137737 · Cited by 231 · see citing works
White, D. A., Pagarette, A., Rooks, P., & Ali, S. T. (2012) · Journal of Applied Phycology · doi:10.1007/s10811-012-9849-6 · Cited by 229 · see citing works
Protein composition of oil bodies in Arabidopsis thaliana ecotype WS
Jolivet, P., Roux, É., d’Andrea, S., et al. (2004) · Plant Physiology and Biochemistry · doi:10.1016/j.plaphy.2004.04.006 · Cited by 218 · see citing works
Visualizing "green oil" in live algal cells
Cooper, M. S., Hardin, W. R., Petersen, T. W., & Cattolico, R. A. (2010) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2009.08.004 · Cited by 215 · see citing works
INFLUENCE OF IRRADIANCE ON THE FATTY ACID COMPOSITION OF PHYTOPLANKTON1
THOMPSON, P. A., Harrison, P. J. H., & Whyte, J. N. C. (1990) · Journal of Phycology · doi:10.1111/j.0022-3646.1990.00278.x · Cited by 205 · see citing works
Biochemical composition of three algal species proposed as food for captive freshwater mussels
Gatenby, C. M., Orcutt, D. M., Kreeger, D. A., Parker, B. C., Jones, V. A., & Neves, R. J. (2003) · Journal of Applied Phycology · doi:10.1023/a:1022929423011 · Cited by 148 · see citing works
Protein and lipid composition analysis of oil bodies from two Brassica napus cultivars
Katavić, V., Agrawal, G. K., Hajdúch, M., et al. (2006) · PROTEOMICS · doi:10.1002/pmic.200600020 · Cited by 145 · see citing works
Cavonius, L. R., Carlsson, N., & Undeland, I. (2014) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-014-8155-3 · Cited by 134 · see citing works
Self-citationMartino, A. D., Bartual, A., Willis, A., et al. (2011) · Protist · doi:10.1016/j.protis.2011.02.002 · Cited by 127 · see citing works
A Lipid Droplet Protein of Nannochloropsis with Functions Partially Analogous to Plant Oleosins
Vieler, A., Brubaker, S. B., Vick, B., & Christoph Benning, A. (2012) · PLANT PHYSIOLOGY · doi:10.1104/pp.111.193029 · Cited by 126 · see citing works
Characterization of major lipid droplet proteins from Dunaliella
Davidi, L., Katz, A., & Pick, U. (2012) · Planta · doi:10.1007/s00425-011-1585-7 · Cited by 106 · see citing works
Acyl lipids of three microalgae
Alonso, D. L., Belarbi, E., Rodríguez‐Ruiz, J., et al. (1998) · Phytochemistry · doi:10.1016/s0031-9422(97)01080-7 · Cited by 86 · see citing works
Selective imaging of saturated and unsaturated lipids by wide-field CARS-microscopy
Heinrich, C. A., Hofer, A., Ritsch, A., et al. (2008) · Optics Express · doi:10.1364/oe.16.002699 · Cited by 74 · see citing works
He, X. N., Allen, J., Black, P. N., Baldacchini, T., Huang, X., Huang, H., Jiang, L., & Lu, Y. F. (2012) · Biomedical Optics Express · doi:10.1364/boe.3.002896 · Cited by 62 · see citing works
De la Hoz Siegler, H., Ayidzoe, W., Ben-Zvi, A., Burrell, R. E., & McCaffrey, W. C. (2012) · Algal Research · doi:10.1016/j.algal.2012.07.004 · Cited by 57 · see citing works
Wong, D. M., & Franz, A. K. (2013) · Journal of Microbiological Methods · doi:10.1016/j.mimet.2013.07.026 · Cited by 45 · see citing works
Coherent Anti-Stokes Raman Scattering Microscopy of Cellular Lipid Storage
Enejder, A. M. K., Brackmann, C., & Svedberg, F. (2010) · IEEE Journal of Selected Topics in Quantum Electronics · doi:10.1109/jstqe.2009.2032512 · Cited by 45 · see citing works
Self-citationErickson, R. A., & Jimenez, R. (2013) · Lab on a Chip · doi:10.1039/c3lc41429a · Cited by 34 · see citing works
Current developments in high‐throughput analysis for microalgae cellular contents
Lee, T.-H., Chang, J., & Wang, H.-Y. A. (2013) · Biotechnology Journal · doi:10.1002/biot.201200391 · Cited by 33 · see citing works
Gorokhova, E., Mattsson, L., & Sundström, A. M. (2012) · Journal of Microbiological Methods · doi:10.1016/j.mimet.2012.03.005 · Cited by 32 · see citing works
Chemical imaging of lignocellulosic biomass by CARS microscopy
Pohling, C., Brackmann, C., Duarte, A. S., Buckup, T., Enejder, A. M. K., & Motzkus, M. (2013) · Journal of Biophotonics · doi:10.1002/jbio.201300052 · Cited by 19 · see citing works
Self-citationFoissner, I. (2009) · PROTOPLASMA · doi:10.1007/s00709-009-0072-5 · Cited by 14 · see citing works
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Raman Spectroscopy and Its Modifications Applied to Biological and Medical Research
Allakhverdiev, E. S., Khabatova, V. V., Kossalbayev, B. D., Zadneprovskaya, E. V., Rodnenkov, O. V., Martynyuk, T. V., Maksimov, G. V., Alwasel, S., Tomo, T., & Allakhverdiev, S. I. (2022) · Cells · doi:10.3390/cells11030386 · Cited by 88 · see citing works
Li, X., Sha, J., Chu, B., Wei, Y., Huang, W., Zhou, H., Xu, N., & He, Y. (2019) · Sensors and Actuators B: Chemical · doi:10.1016/j.snb.2019.04.048 · Cited by 52 · see citing works
Jaeger, D., Pilger, C., Hachmeister, H., Oberländer, E., Wördenweber, R., Wichmann, J., Mussgnug, J. H., Huser, T., & Kruse, O. (2016) · Scientific Reports · doi:10.1038/srep35340 · Cited by 47 · see citing works
- Authors
- Cavonius, L., Fink, H., Kiskis, J., Albers, E., Undeland, I., & Enejder, A.
- Affiliations
- Chalmers University of Technology (SE)
- Division of Life Science, Department of Chemical and Biological Engineering, Chalmers University of Technology, SE–412 96 Gothenburg, Sweden
- Venue
- Plant Physiology
- Publisher
- Oxford University Press (OUP)
- Published
- 2015-01-12
- Volume
- 167
- Issue
- 3
- Pages
- 603-616
- ISSN
- 0032-0889 (ISSN-L)
- 1532-2548
- PMC
- PMC4348760
- OpenAlex
- W2107107529
- Cited by
- 44
- References
- 41
- Citations by year
- 2025: 7 · 2024: 6 · 2023: 1 · 2022: 2 · 2021: 6 · 2020: 1 · 2019: 4 · 2018: 5 · 2017: 5 · 2016: 6 · 2015: 1
- FWCI
- 3.43
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- PLANT PHYSIOLOGY (publishedVersion, cc-by)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Chalmers Research (Chalmers University of Technology) (submittedVersion)
- Chalmers Publication Library (Chalmers University of Technology) (submittedVersion)
- Funders
- Chalmers Tekniska Högskola
- Vetenskapsrådet
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
- Keywords
- microalgae, lipid droplets, coherent anti-Stokes Raman scattering microscopy, Phaeodactylum tricornutum, lipid accumulation, nitrogen starvation, light limitation, lipid droplet fusion, lipid fluidity, fatty acid saturation, autofluorescence, biodiesel, nutraceuticals
- MeSH terms
- Cell Survival, Cells, Cultured, Fatty Acids, Lipids, Gas Chromatography-Mass Spectrometry, Membrane Fluidity, Microscopy, Spectrum Analysis, Raman, Time Factors, Imaging, Three-Dimensional, Intracellular Space, Microalgae, Lipid Droplets
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