Alberto Pérez-Huerta

Multiscale structure of calcite fibres of the shell of the brachiopod Terebratulina retusa

The shells of rhynchonelliform brachiopods have an outer (primary) layer of acicular calcite and an inner (secondary) layer of calcite fibres which are parallel to the shell exterior. Atomic force microscopy (AFM) reveals that these fibres are composed of large triangular nanogranules of about 600-650 nm along their long axis. The nanogranules are composites of organic and inorganic components. As the shell grows, the fibres elongate with the calcite c-axis perpendicular to the fibre axis as demonstrated by electron backscatter diffraction (EBSD). Thus, despite being a composite structure comprising granules that are themselves composites, each fibre is effectively a single crystal. The combination of AFM and EBSD reveals the details of the structure and crystallography of these fibres. This knowledge serves to identify those aspects of biological control that must be understood to enable comprehension of the biological control exerted on the construction of these exquisite biomineral structures.

Material properties of brachiopod shell ultrastructure by nanoindentation

Mineral-producing organisms exert exquisite control on all aspects of biomineral production. Among shell-bearing organisms, a wide range of mineral fabrics are developed reflecting diverse modes of life that require different material properties. Our knowledge of how biomineral structures relate to material properties is still limited because it requires the determination of these properties on a detailed scale. Nanoindentation, mostly applied in engineering and materials science, is used here to assess, at the microstructural level, material properties of two calcite brachiopods living in the same environment but with different modes of life and shell ultrastructure. Values of hardness (H) and the Young modulus of elasticity (E) are determined by nanoindentation. In brachiopod shells, calcite semi-nacre provides a harder and stiffer structure (H∼3–6 GPa; E=60–110/120 GPa) than calcite fibres (H=0–3 GPa; E=20–60/80 GPa). Thus, brachiopods with calcite semi-nacre can cement to a substrate and remain immobile during their adult life cycle. This correlation between mode of life and material properties, as a consequence of ultrastructure, begins to explain why organisms produce a wide range of structures using the same chemical components, such as calcium carbonate.

Optimizing Electron Backscatter Diffraction of Carbonate Biominerals—Resin Type and Carbon Coating

Electron backscatter diffraction (EBSD) is becoming a widely used technique to determine crystallographic orientation in biogenic carbonates. Despite this use, there is little information available on preparation for the analysis of biogenic carbonates. EBSD data are compared for biogenic aragonite and calcite in the common blue mussel, Mytilus edulis, using different types of resin and thicknesses of carbon coating. Results indicate that carbonate biomineral samples provide better EBSD results if they are embedded in resin, particularly epoxy resin. A uniform layer of carbon of 2.5 nm thickness provides sufficient conductivity for EBSD analyses of such insulators to avoid charging without masking the diffracted signal. Diffraction intensity decreases with carbon coating thickness of 5 nm or more. This study demonstrates the importance of optimizing sample preparation for EBSD analyses of insulators such as carbonate biominerals.

Oxygen isotope composition in Modiolus modiolus aragonite in the context of biological and crystallographic control

Abstract Living systems exert exquisite control on all aspects of biomineral production and organic components, including proteins, are essential to this biological control. The protein-rich extrapallial (EP) fluid of bivalve molluscs is a strong candidate for the source of such proteins. Differences in calcium carbonate polymorphs between Modiolus modiolus and Mytilus edulis are concurrent with differences in EP fluid protein profiles. In conjunction with this biological control is the environmental influence which is interpreted using proxies such as δ18O to determine the history of ambient seawater temperature. In the horse mussel, Modiolus modiolus, the difference in oxygen isotope fractionation in the nacreous aragonite and the prismatic aragonite layer results in respective δ18O values of 2.1±0.2% and 2.5±0.2‰. These δ18O values result in estimates of ambient seawater of 12.1±0.6°C and 10.2±0.6°C for nacreous and prismatic aragonite, respectively. Electron backscatter diffraction is used here to determine the crystallographic orientation at high spatial resolution, allowing the measurements of stable isotopes to be accurately mapped in terms of shell architecture. These preliminary data suggest that it is essential to account for both polymorph and crystal habit when deciphering ambient seawater temperature using δ18O as a proxy.

Structural, Mineralogical, and Biochemical Diversity in the Lower Part of the Pearl Layer of Cultivated Seawater Pearls from Polynesia

A series of Polynesian pearls has been investigated with particular attention to the structural and compositional patterns of the early developmental stages of the pearl layer. These initial steps in pearl formation bear witness of the metabolic changes that have occurred during the pearl-sac formation. The resulting structurally and biochemically complex structures have been investigated using a variety of techniques that provide us with information concerning both mineral phases and the organic components. Results are discussed with respect to our understanding of the biomineralization mechanisms, as well as for the grafting process.

Deciphering the distribution of organic components in brachiopod shells by confocal laser scanning microscopy

Characterization of the nature and distribution of organic components is crucial to understand shell formation in marine invertebrates. Although several techniques can provide detailed information at high spatial resolution, few of them are non‐destructive and informative in a larger structural context. We explore the use of confocal laser scanning microscopy (CLSM) to obtain a better understanding of the distribution of organic components in calcitic shells of brachiopods focusing on perforations (punctae) across the shell. Resulting intensities and patterns of fluorescence correspond well with the distribution of polysaccharides and proteins as reported in previous histological and biochemical studies. Confocal laser microscopy is, therefore, a useful tool to be combined with other techniques to improve our knowledge of biomineral structures in marine invertebrates.

Assessment of crystallographic influence on material properties of calcite brachiopods

Abstract Calcium carbonate biominerals are frequently analysed in materials science due to their abundance, diversity and unique material properties. Aragonite nacre is intensively studied, but less information is available about the material properties of biogenic calcite, despite its occurrence in a wide range of structures in different organisms. In particular, there is insufficient knowledge about how preferential crystallographic orientations influence these material properties. Here, we study the influence of crystallography on material properties in calcite semi-nacre and fibres of brachiopod shells using nano-indentation and electron backscatter diffraction (EBSD). The nano-indentation results show that calcite semi-nacre is a harder and stiffer (H ≈ 3-5 GPa; E = 50-85 GPa) biomineral structure than calcite fibres (H = 0.4-3 GPa; E = 30-60 GPa). The integration of EBSD to these studies has revealed a relationship between the crystallography and material properties at high spatial resolution for calcite semi-nacre. The presence of crystals with the c-axis perpendicular to the plane-of-view in longitudinal section increases hardness and stiffness. The present study determines how nano-indentation and EBSD can be combined to provide a detailed understanding of biomineral structures and their analysis for application in materials science.

Influence of crystallographic orientation of biogenic calcite on in situ Mg XANES analyses

Micro X-ray absorption near-edge spectroscopy at the Mg K-edge is a useful technique for acquiring information about the environment of Mg(2+) in biogenic calcite. These analyses can be applied to shell powders or intact shell structures. The advantage of the latter is that the XANES analyses can be applied to specific areas, at high (e.g. micrometre) spatial resolution, to determine the environment of Mg(2+) in a biomineral context. Such in situ synchrotron analysis has to take into account the potential effect of crystallographic orientation given the anisotropy of calcite crystals and the polarized nature of X-rays. Brachiopod shells of species with different crystallographic orientations are used to assess this crystallographic effect on in situ synchrotron measurements at the Mg K-edge. Results show that, owing to the anisotropy of calcite, in situ X-ray absorption spectra (XAS) are influenced by the crystallographic orientation of calcite crystals with a subsequent potentially erroneous interpretation of Mg(2+) data. Thus, this study demonstrates the importance of crystallography for XAS analyses and, therefore, the necessity to obtain crystallographic information at high spatial resolution prior to spectroscopic analysis.

Crystallography of craniid brachiopods by electron backscatter diffraction (EBSD)

Electron backscatter diffraction (EBSD) is used to determine the detailed crystallographic orientation of calcite crystals of craniid brachiopods in the context of shell ultrastructure. Sections of shells of two Recent species, Novocrania anomala and Novocrania huttoni , are analysed to provide 3D crystallographic patterns at high spatial resolution. The c -axis of semi-nacre calcite crystals is oriented parallel to the laminae that define the ultrastructure of the secondary layer. This orientation differs from that of rhynchonelliform calcitic brachiopods where the c -axis is perpendicular to the length of morphological fibres and to the shell exterior.

Oxygen isotope equilibrium in brachiopod shell fibres in the context of biological control

With their long geological history and stable low-Mg calcite shells, Rhynchonelliform brachiopods are attractive sources of environmental data such as past seawater temperature (Buening and Spero, 1996; Auclair et al. , 2003; Brand et al. , 2003; Parkinson et al. , 2005). Concerns about the influence of vital effects on the stable isotope composition of brachiopod shells (Popp et al. , 1986), led to isotope analyses of different parts of brachiopod shells in order to identify those parts of the shell that are influenced by any vital effect and those parts that may be suitable recorders of seawater temperature via stable oxygen isotope composition (Carpenter and Lohmann, 1995; Parkinson et al. , 2005). Such detailed studies demonstrated that the outer primary layer of acicular calcite is isotopically light in both δ18O and δ13C while the secondary layer, composed of calcite fibres, is in oxygen-isotope equilibrium with ambient seawater (Fig. 1⇓) (Parkinson et al. , 2005). Use of the isotopically-light primary layer calcite in temperature calculations would result in seawater temperatures that are significantly higher than the actual ambient seawater temperature (Parkinson et al. , 2005). Isotopic equilibrium between ambient seawater and …