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In vivo and in situ synchrotron radiation-based μ-XRF reveals elemental distributions during the early attachment phase of barnacle larvae and juvenile barnacles

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Abstract

Barnacles are able to establish stable surface contacts and adhere underwater. While the composition of adult barnacle cement has been intensively studied, far less is known about the composition of the cement of the settlement-stage cypris larva. The main challenge in studying the adhesives used by these larvae is the small quantity of material available for analysis, being on the order of nanograms. In this work, we applied, for the first time, synchrotron radiation-based μ-X-ray fluorescence analysis (SR-μ-XRF) for in vivo and in situ analysis of young barnacles and barnacle cyprids. To obtain biologically relevant information relating to the body tissues, adhesives, and shell of the organisms, an in situ sample environment was developed to allow direct microprobe investigation of hydrated specimens without pretreatment of the samples. In 8-day-old juvenile barnacles (Balanus improvisus), the junctions between the six plates forming the shell wall showed elevated concentrations of calcium, potassium, bromine, strontium, and manganese. Confocal measurements allowed elemental characterization of the adhesive interface of recently attached cyprids (Balanus amphitrite), and substantiated the accumulation of bromine both at the point of initial attachment as well as within the cyprid carapace. In situ measurements of the cyprid cement established the presence of bromine, chlorine, iodine, sulfur, copper, iron, zinc, selenium, and nickel for both species. The previously unrecognized presence of bromine, iron, and selenium in the cyprid permanent adhesive will hopefully inspire further biochemical investigations of the function of these substances.

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Acknowledgments

The authors acknowledge the Synchrotron Light Source ANKA for provision of beamtime at the FLUO beamline and the ANKA staff for excellent support during the beamtime. This work was funded by the Virtual Institute VH-VI-403 of the Helmholtz Association, Office of Naval Research grants N00014-12-1-0498 and N00014-15-1-2324, and the 7th framework International Marie-Curie Training network SeaCoat (grant 237997). N. Aldred and A. S. Clare acknowledge funding support from Office of Naval Research grants N00014-08-1-1240 to A.S.C. and N00014-13-1-0633 to A.S.C. and N.A.

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Authors and Affiliations

  1. Institute of Functional Interfaces, KIT Karlsruhe Institute of Technology, Hermann-von-Helmholtz-Platz 1, 76344, Eggenstein-Leopoldshafen, Germany

    Tobias Senkbeil & Tawheed Mohamed

  2. Analytical Chemistry - Biointerfaces, Ruhr-University Bochum, Universitätsstr. 150 NC4, 44801, Bochum, Germany

    Tobias Senkbeil & Axel Rosenhahn

  3. Applied Physical Chemistry, University of Heidelberg, Im Neuenheimer Feld 253, 69120, Heidelberg, Germany

    Tawheed Mohamed

  4. Institute of Physics and Technology, International X-ray Optics Lab, National Research Tomsk Polytechnic University (TPU), 30, Lenin ave., Tomsk, 634050, Russia

    Tawheed Mohamed

  5. ANKA Synchrotronstrahlungsquelle, KIT Karlsruhe Institute of Technology, Hermann-von-Helmholtz-Platz 1, 76344, Eggenstein-Leopoldshafen, Germany

    Rolf Simon & David Batchelor

  6. School of Marine Science and Technology, Newcastle University, Newcastle upon Tyne, NE1 7RU, UK

    Alessio Di Fino, Nick Aldred & Anthony S. Clare

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  1. Tobias Senkbeil
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Correspondence to Axel Rosenhahn.

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Senkbeil, T., Mohamed, T., Simon, R. et al. In vivo and in situ synchrotron radiation-based μ-XRF reveals elemental distributions during the early attachment phase of barnacle larvae and juvenile barnacles. Anal Bioanal Chem 408, 1487–1496 (2016). https://doi.org/10.1007/s00216-015-9253-6

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  • DOI: https://doi.org/10.1007/s00216-015-9253-6

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