Callow, J.A.

Callow, M.E.

Grunze, M.

Heydt, M.

Rosenhahn, A.

Stuppy, S.

Swain, G.

English

Callow, J. A.

Callow, M. E.

KITopen

KIT – University of the State of Baden-Wuerttemberg andNational Research Center of the Helmholtz AssociationInvestigation of motile marine organisms with Digital In-line Holographic Microscopy in situ and under lab conditionsSvenja Stuppy1,3*, Matthias Heydt1,Geoffrey W. Swain4, J.A. Callow5, M.E. Callow5, Michael Grunze1,2, Axel Rosenhahn1,31 Applied Physical Chemistry, Ruprecht-Karls-University Heidelberg, Im Neuenheimer Feld 253, 69120 Heidelberg, Germany2 Institute for Toxicology and Genetics, ITG, Karlsruhe Institute of Technology,PO Box 3640, 76021 Karlsruhe, Germany3 Institute for Functional Interfaces, IFG, Karlsruhe Institute of Technology,PO Box 3640, 76021 Karlsruhe, Germany4 Ocean Engineering and Oceanography, Florida Institute of Technology, 150 West University Boulevard, Melbourne Florida 32901, USA5 School of Bioscience, University of Birmingham, Birmingham B15 2TT, UKAcknowledgment• Thanks to the group of Prof. Swain Florida Institute of Technology• The work was funded by Office of Naval Research(Grant number N00014-08-1-1116) and DFG 252412-2Introduction Biofouling caused by different micro- and macro-organisms such as algae, diatoms or invertebrates is a major problem in marine industries [1]. The investigation of the settlement behavior of these organisms on different surfaces or coatings allows to reveal selection strategies.In previous work swimming and settlement behavior of Ulva linza zoospores under lab conditions was observed in the vicinity of different chemistries with a digital in-line holographic Microscope [2].For the investigation of marine organisms in their native environment experiments on a test facility situated at the Indian River Lagoon in Melbourne Beach Florida were performed. Field experimentsHolographyReconstructionResolution L2DNA =Hologram 3D-ProjectionKirschhoff-Helmholtz Transformation• Transportable holographic Setup in in-line geometry, first proposedby Gabor in 1948 [3]• Experiments performed in a mobile laboratory (working in a Van)• independent from electrical power supply; working with car batteries• Seawater taken out of seawater tank and filtered with 30µm netfilters•Record of Hologram series with up to 18000 frames (10 frames per second )Detector (CMOS Chip)DPSS Laser 532 nmAperturesBeamexpander 20x ObjectivSample translationstage500nm PinholeMirrorsSource Hologram Twin image Self interference2scscinscin2in )()()()()()()( rrrrrrrIΨ+ΨΨ+ΨΨ+Ψ= ∗∗ξd ξrξikexp )ξI()rK(s2∫ =NA0,61λδlateral = 2depth NAλδ =Tracking Marine Biofoulers71.75 284.8Ulva Linza spore on AWGOrientationpatternSetupThe motile zoospores of the green algae Ulva linza areestablished model organisms to investigate explorationbehaviour and settlement dynamics related to different chemicalcues. Settlement is a critical stage in Ulva life cycle. Theselection of a suitable surface is important for their survival andreproduction. However Ulva linza has to be collected from aseashore prior to experiments. The zoospores can be releasedand measured in the laboratory at University of Birmingham.The step from a model organism to a conglomeration of marinebiofoulers under native conditions is a big challenge butnecessary to correlate between lab and field.Comparison Lab versus FieldLab [4]FieldMean velocity of tracked organismsRound organismOrientationpatternField Lab LabMarine organisms after 3 h in observation chamberUlva spores after 35 s in observation chamberUlva spores after 11:57 min in observation chamberLiterature: [1] Rosenhahn, A., Ederth, T. & Pettit, M. E. Advanced Nanostructures for the control of biofouling: The FP6 EU Integrated Project AMBIO. Biointerphases 3, IR1-IR5 (2008)[2] Heydt, M. et al. Digital in-line holography as a three-dimensional tool to study motile marine organisms during their exploration of surfaces. Journal of Adhesion83,417-430 (2007)[3] Gabor, D. (1948). "A new microscopic principle." Nature 161: 777-778[4] Matthias Heydt How do spores select where to settle? Springer Theses ISBN 978-3-642-17216-8OutlookObservation of active swimming organisms with elongated shapeVelocity coded 3D-trajectoriesVelocity distributions of Ulva linza on AWG and spherical undefinedorganisms measured in the field•Velocity of Ulva linza spores under lab conditions shows a split in a slow and a fast swimming fraction after longerincubation time in the observation chamber•Organisms in field experiment show a broader velocitydistribution without spliting even after a very long incubationtime•Velocity distribution of organisms with elongated shapesplits in two fractionsfast swimmers (mean velocity: 284.8 µm/s) and slowswimmers (mean velocity:71.75 µm/s)•Elongated organisms swim along thier longitudinal axisand have a round shape relating to their transversal axis3D trajectory with shape of organismsobserved in xy-projections•Reproduction and further investigations of settlement behaviour of Ulva linza spores on surfaces withdifferent surface properties•Differentiation and possibly identification of marine species from field experiments on the basis of theirshape and velocity•Further correlation of field work with lab experiments•Correlation of holographic data with other data obtained in the field (settlement kinetics,  Microfluidics, Conditioning film formation, ect.)Organism with elongatedshapeSpherical organismxy-projection*email: svenja.stuppy@pci.uni-heidelberg.de

Investigation of motile marine organisms with digital in-line holographic microscopy in situ and under lab conditions

https://publikationen.bibliothek.kit.edu/220085831/3815964

Investigation of motile marine organisms with digital in-line holographic microscopy in situ and under lab conditions

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