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The use of dual beam ESEM FIB to reveal the internal ultrastructure of hydroxyapatite nanoparticle-sugar-glass composites

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Abstract

Microparticles (MP) spray dried from hydroxyapatite (HA) nanoparticle (NP) sugar suspensions are currently under development as a prolonged release vaccine vehicle. Those with a significant sugar component cannot be sectioned by ultramicrotomy as resins are excluded by the sugar. Focused ion beam (FIB) milling is the only method to prepare thin sections that enables the inspection of the MPs ultrastructure by transmission electron microscopy (TEM). Several methods have been explored and we have found it is simplest to encapsulate MPs in silver dag, sandwiched between gold foils for FIB-milling to enable multiple MPs to be sectioned simultaneously. Spray dried MPs containing 80% sugar have an inter-nanoparticle separation that is comparable with NP size (~50 nm). MPs spray dried with 50% sugar or no sugar are more tightly packed. Nano-porosity in the order of 10 nm exists between NPs. MPs spray dried in the absence of sugar and sectioned by ultramicrotomy or by FIB-milling have comparable nanoscale morphologies. Selected area electron diffraction (SAED) demonstrates that the HA remains (substantially) crystalline following FIB-milling.

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Acknowledgments

This work was funded by a DTI grant (CHBS/004/00063C) held jointly by J. N. Skepper at the University of Cambridge and Cambridge Biostability Ltd. The Dual Beam FIB/ESEM was purchased with funds from the BBSRC.

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Authors and Affiliations

  1. Multi-Imaging Centre, Department of Physiology, Development and Neuroscience, School of Biological Sciences, University of Cambridge, Anatomy Building, Downing Street, Cambridge, CB2 3DY, UK

    David M. Wright, Michael Motskin & Jeremy N. Skepper

  2. Department of Physics, Cavendish Laboratory, University of Cambridge, J J Thomson Avenue, Cambridge, CB3 0HE, UK

    John J. Rickard, Harald Dobberstein & Athene M. Donald

  3. Cambridge Biostability Ltd., Unit 184, Cambridge Science Park, Milton Road, Cambridge, Cambridgeshire, CB4 0GA, UK

    Nigel H. Kyle & Tevor G. Gard

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  1. David M. Wright
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Correspondence to David M. Wright.

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Wright, D.M., Rickard, J.J., Kyle, N.H. et al. The use of dual beam ESEM FIB to reveal the internal ultrastructure of hydroxyapatite nanoparticle-sugar-glass composites. J Mater Sci: Mater Med 20, 203–214 (2009). https://doi.org/10.1007/s10856-008-3539-6

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