Detachment of Dunaliella tertiolecta Microalgae from a Glass Surface by a Near-Infrared Optical Trap

We report on the observation of the detachment in situ and in vivo of Dunaliella tertiolecta microalgae cells from a glass surface using a 1064 nm wavelength trapping laser beam. The principal bends of both flagella of Dunaliella were seen self-adhered to either the top or bottom coverslip surfaces of a 50 μm thick chamber. When a selected attached Dunaliella was placed in the trapping site, it photoresponded to the laser beam by moving its body and flagellar tips, which eventually resulted in its detachment. The dependence of the time required for detachment on the trapping power was measured. No significant difference was found in the detachment time for cells detached from the top or bottom coverslip, indicating that the induced detachment was not due solely to the optical forces applied to the cells. After detachment, the cells remained within the optical trap. Dunaliella detached from the bottom were seen rotating about their long axis in a counterclockwise direction, while those detached from the top did not rotate. The rotation frequency and the minimal force required to escape from the trap were also measured. The average rotation frequency was found to be independent of the trapping power, and the swimming force of a cell escaping the laser trap ranged from 4 to 10 picoNewtons. Our observations provide insight into the photostimulus produced when a near-infrared trapping beam encounters a Dunaliella. The microalgae frequently absorb more light than they can actually use in photosynthesis, which could cause genetic and molecular changes. Our findings may open new research directions into the study of photomovement in species of Dunaliella and other swimming microorganisms that could eventually help to solve technological problems currently confronting biomass production. In future work, studies of the response to excess light may uncover unrecognized mechanisms of photoprotection and photoacclimation.

[1]  N. J. Antia,et al.  ULTRASTRUCTURAL OBSERVATION OF THE SURFACE COAT OF DUNALIELLA TERTIOLECTA FROM STAINING WITH CATIONIC DYES AND ENZYME TREATMENTS , 1980 .

[2]  Aharon Oren,et al.  THE ALGA DUNALIELLA. BIODIVERSITY, PHYSIOLOGY, GENOMICS AND BIOTECHNOLOGY , 2010 .

[3]  William H. Guilford,et al.  Laser Trap Characterization and Modeling of Phototaxis in Chlamydomonas reinhardtii , 2009 .

[4]  R. Uhl,et al.  On the localization of voltage-sensitive calcium channels in the flagella of Chlamydomonas reinhardtii , 1994, The Journal of cell biology.

[5]  Mario Montes-Usategui,et al.  Influence of experimental parameters on the laser heating of an optical trap , 2017, Scientific Reports.

[6]  A. Ashkin,et al.  Optical trapping and manipulation of single cells using infrared laser beams , 1987, Nature.

[7]  Veneranda G. Garces,et al.  Induced deflagellation of Isochrysis microalgae in a near-infrared optical trap , 2015 .

[8]  U. Rüffer,et al.  Comparison of the beating of cis‐ and trans‐flagella of Chlamydomonas cells held on micropipettes , 1987 .

[9]  Gabriel C. Spalding,et al.  Motility assessment of green biflagellated microalgae in an optical trap using back focal plane interferometry , 2019, NanoScience + Engineering.

[10]  B. Rao,et al.  Flagella-generated forces reveal gear-type motor in single cells of the green alga, Chlamydomonas reinhardtii. , 2009, Biochemical and biophysical research communications.

[11]  Monika Ritsch-Marte,et al.  Optical macro-tweezers: trapping of highly motile micro-organisms , 2011 .

[12]  J. Rosenbaum,et al.  Intraflagellar transport , 2002, Nature Reviews Molecular Cell Biology.

[13]  M. Schliwa,et al.  Calibration of light forces in optical tweezers. , 1995, Applied optics.

[14]  G. Pazour,et al.  The Chlamydomonas Flagellum as a Model for Human Ciliary Disease , 2009 .

[15]  Linda Z. Shi,et al.  The use of optical tweezers to study sperm competition and motility in primates , 2008, Journal of The Royal Society Interface.

[16]  Byung-Kwan Cho,et al.  Effects of Light Intensity and Nitrogen Starvation on Glycerolipid, Glycerophospholipid, and Carotenoid Composition in Dunaliella tertiolecta Culture , 2013, PloS one.

[17]  Raymond E. Goldstein,et al.  Ciliary contact interactions dominate surface scattering of swimming eukaryotes , 2013, Proceedings of the National Academy of Sciences.

[18]  Robert A. Bloodgood The future of ciliary and flagellar membrane research , 2012, Molecular biology of the cell.

[19]  Erik Schäffer,et al.  Under-filling trapping objectives optimizes the use of the available laser power in optical tweezers. , 2011, Optics express.

[20]  M. Bessen,et al.  Calcium control of waveform in isolated flagellar axonemes of chlamydomonas , 1980, The Journal of cell biology.

[21]  D. Mitchell Orientation of the central pair complex during flagellar bend formation in Chlamydomonas. , 2003, Cell motility and the cytoskeleton.

[22]  K. Foster,et al.  Analysis of the ciliary/flagellar beating of Chlamydomonas. , 2009, Methods in cell biology.

[23]  P. Zemánek,et al.  Optical trapping of microalgae at 735-1064 nm: photodamage assessment. , 2013, Journal of photochemistry and photobiology. B, Biology.

[24]  J. Barber,et al.  beta-Carotene quenches singlet oxygen formed by isolated photosystem II reaction centers. , 1994, Biochemistry.

[25]  P. A. Luque,et al.  Effects of the water-soluble fraction of the mixture fuel oil/diesel on the microalgae Dunaliella tertiolecta through growth , 2020, Environmental Science and Pollution Research.

[26]  Gabriel C. Spalding,et al.  Studies of biflagellated microalgae adhesion using an optical trap system , 2018, NanoScience + Engineering.

[27]  C. Lindemann,et al.  Flagellar and ciliary beating: the proven and the possible , 2010, Journal of Cell Science.

[28]  H. Busscher,et al.  Specific and non-specific interactions in bacterial adhesion to solid substrata , 1987 .

[29]  D. L. Ringo FLAGELLAR MOTION AND FINE STRUCTURE OF THE FLAGELLAR APPARATUS IN CHLAMYDOMONAS , 1967, The Journal of cell biology.

[30]  J. W. G. Lund,et al.  An Introductory Account of the Smaller Algae of British Coastal Waters. Part V. Bacillariophyceae (Diatoms). , 1965 .

[31]  W. Marshall,et al.  Total internal reflection fluorescence (TIRF) microscopy of Chlamydomonas flagella. , 2009, Methods in cell biology.

[32]  K. Niyogi,et al.  Sensing and responding to excess light. , 2009, Annual review of plant biology.

[33]  Steven M. Block,et al.  Optical tweezers : a new tool for biophysics , 1990 .

[34]  Johannes Buder Zur Kenntnis der phototaktischen Richtungsbewegungen , 1917 .

[35]  W. J. V. Osterhout,et al.  ON THE DYNAMICS OF PHOTOSYNTHESIS , 1918, The Journal of general physiology.

[36]  C. Linne,et al.  Adhesion of Chlamydomonas microalgae to surfaces is switchable by light , 2017, Nature Physics.

[37]  O. Sineshchekov,et al.  Photoreceptor electric potential in the phototaxis of the alga Haematococcus pluvialis , 1978, Nature.

[38]  L. Y. Maluf,et al.  Ultrastructure of the green flagellate Dunaliella tertiolecta (Chlorophyceae, Volvocales) with comparative notes on three other species , 1981 .

[39]  Paul Stoodley,et al.  Bacterial biofilms: from the Natural environment to infectious diseases , 2004, Nature Reviews Microbiology.

[40]  B. Rao,et al.  Sensitive, real-time monitoring of UV-induced stress in a single, live plant cell using an optical trap , 2006 .

[41]  Y. Posudin,et al.  Photomovement of Dunaliella Teod. , 2010 .

[42]  D Mathur,et al.  Optically-controllable, micron-sized motor based on live cells. , 2005, Optics express.

[43]  M. Shariati,et al.  Dunaliella biotechnology: methods and applications , 2009, Journal of applied microbiology.

[44]  M. Couturier,et al.  Ciliary beat and cell motility of Dunaliella: computer analysis of high speed microcinematography , 1988 .

[45]  G. B. Witman,et al.  CHLAMYDOMONAS FLAGELLA , 1972, The Journal of cell biology.

[46]  M. Kornaros,et al.  Effects of Burkholderia thailandensis rhamnolipids on the unicellular algae Dunaliella tertiolecta. , 2019, Ecotoxicology and environmental safety.

[47]  D. Singh,et al.  A re view on pharmacological applications of halophilic alga Dunaliella , 2016 .

[48]  D. Arnon Sunlight, Earth Life , 1982 .

[49]  U. Rüffer,et al.  Flagellar photoresponses ofChlamydomonascells held on micropipettes: II. Change in flagellar beat pattern: Flagellar Beat Pattern Change inchlamydomonas , 1991 .

[50]  B. Grodzinski,et al.  Energy balance, organellar redox status, and acclimation to environmental stress , 2006 .

[51]  E. Molina-Grima,et al.  Biofouling in photobioreactors for marine microalgae , 2017, Critical reviews in biotechnology.

[52]  R. McCord,et al.  Analysis of force generation during flagellar assembly through optical trapping of free-swimming Chlamydomonas reinhardtii. , 2005, Cell motility and the cytoskeleton.

[53]  A. Oren A hundred years of Dunaliella research: 1905–2005 , 2005, Saline systems.

[54]  K. Niyogi,et al.  PHOTOPROTECTION REVISITED: Genetic and Molecular Approaches. , 1999, Annual review of plant physiology and plant molecular biology.

[55]  Enkeleida Lushi,et al.  Microalgae Scatter off Solid Surfaces by Hydrodynamic and Contact Forces. , 2015, Physical review letters.

[56]  U. Rüffer,et al.  Flagellar photoresponses of Chlamydomonas cells held on micropipettes: II. Change in flagellar beat pattern , 1990 .

[57]  Idan Tuval,et al.  Noise and synchronization in pairs of beating eukaryotic flagella. , 2009, Physical review letters.

[58]  A. Ashkin,et al.  Optical trapping and manipulation of viruses and bacteria. , 1987, Science.

[59]  G. Sonek,et al.  Evidence for localized cell heating induced by infrared optical tweezers. , 1995, Biophysical journal.

[60]  A. Krieger-Liszkay Singlet oxygen production in photosynthesis. , 2004, Journal of experimental botany.