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Instruments / Technical Area	Publication Details	Published
I13-2-Diamond Manchester Imaging	Imaging techniques for observing laminar geometry in the feather shaft cortex Christian Laurent , Sharif Ahmed , Richard Boardman , Richard Cook , Gareth Dyke , Colin Palmer , Philipp Schneider , Roland De Kat Journal Of Microscopy DOI: 10.1111/jmi.12820 Show Abstract ▼ Abstract: Bird feather shafts are light, stiff, and strong, but the fine details of how their structure, mechanics and function relate to one another remains poorly understood. The missing piece in our understanding may be the various fibrous layers that make up the shaft's cortex. Detailed imaging techniques are needed to enable us to capture, analyse and quantify these layers before we can begin to unravel the relationship between their structure, mechanics and function. We show that Serial‐Block‐Face scanning electron microscopy, scanning confocal polarised microscopy and synchrotron‐based computed tomography are three suitable techniques to investigate layer thickness and fibre orientation in the feather cortex. These techniques and other are discussed in terms of their ability to resolve the fibrous laminar structure of the feather cortex, on sample preparation, and on throughput. Annotated images are presented for each and less suitable techniques are presented in the supplement.	Jun 2019
I13-2-Diamond Manchester Imaging	Optimization of digital volume correlation computation in SR-microCT images of trabecular bone and bone-biomaterial systems M. Peña Fernández , A. H. Barber , G. W. Blunn , G. Tozzi Journal Of Microscopy 373 DOI: 10.1111/jmi.12745 Diamond Proposal Number(s): [14080] Show Abstract ▼ Abstract: A micromechanical characterization of biomaterials for bone tissue engineering is essential to understand the quality of the newly regenerated bone, enabling the improvement of tissue regeneration strategies. A combination of microcomputed tomography in conjunction with in situ mechanical testing and digital volume correlation (DVC) has become a powerful technique to investigate the internal deformation of bone structure at a range of dimensional scales. However, in order to obtain accurate three‐dimensional strain measurement at tissue level, high‐resolution images must be acquired, and displacement/strain measurement uncertainties evaluated. The aim of this study was to optimize imaging parameters, image postprocessing and DVC settings to enhance computation based on ‘zero‐strain’ repeated high‐resolution synchrotron microCT scans of trabecular bone and bone‐biomaterial systems. Low exposures to SR X‐ray radiation were required to minimize irradiation‐induced tissue damage, resulting in the need of advanced three‐dimensional filters on the reconstructed images to reduce DVC‐measured strain errors. Furthermore, the computation of strain values only in the hard phase (i.e. bone, biomaterial) allowed the exclusion of large artefacts localized in the bone marrow. This study demonstrated the suitability of a local DVC approach based on synchrotron microCT images to investigate the micromechanics of trabecular bone and bone‐biomaterial composites at tissue level with a standard deviation of the errors in the region of 100 microstrain after a thorough optimization of DVC computation.	Jul 2018
I13-2-Diamond Manchester Imaging	A review of techniques for visualising soft tissue microstructure deformation and quantifying strain Ex Vivo C. M. Disney , P. D. Lee , J. A. Hoyland , M. J. Sherratt , B. K. Bay Journal Of Microscopy 35 DOI: 10.1111/jmi.12701 Diamond Proposal Number(s): [12776] Show Abstract ▼ Abstract: Many biological tissues have a complex hierarchical structure allowing them to function under demanding physiological loading conditions. Structural changes caused by ageing or disease can lead to loss of mechanical function. Therefore, it is necessary to characterise tissue structure to understand normal tissue function and the progression of disease. Ideally intact native tissues should be imaged in 3D and under physiological loading conditions. The current published in situ imaging methodologies demonstrate a compromise between imaging limitations and maintaining the samples native mechanical function. This review gives an overview of in situ imaging techniques used to visualise microstructural deformation of soft tissue, including three case studies of different tissues (tendon, intervertebral disc and artery). Some of the imaging techniques restricted analysis to observational mechanics or discrete strain measurement from invasive markers. Full‐field local surface strain measurement has been achieved using digital image correlation. Volumetric strain fields have successfully been quantified from in situ X‐ray microtomography (micro‐CT) studies of bone using digital volume correlation but not in soft tissue due to low X‐ray transmission contrast. With the latest developments in micro‐CT showing in‐line phase contrast capability to resolve native soft tissue microstructure, there is potential for future soft tissue mechanics research where 3D local strain can be quantified. These methods will provide information on the local 3D micromechanical environment experienced by cells in healthy, aged and diseased tissues. It is hoped that future applications of in situ imaging techniques will impact positively on the design and testing of potential tissue replacements or regenerative therapies.	Apr 2018
	An optical configuration for fastidious STEM detector calibration and the effect of the objective-lens pre-field L. Jones , A. Varambhia , H. Sawada , P. D. Nellist Journal Of Microscopy 180 DOI: 10.1111/jmi.12672 Show Abstract ▼ Abstract: In the scanning transmission electron microscope, an accurate knowledge of detector collection angles is paramount in order to quantify signals on an absolute scale. Here we present an optical configuration designed for the accurate measurement of collection angles for both image-detectors and energy-loss spectrometers. By deflecting a parallel electron beam, carefully calibrated using a diffraction pattern from a known material, we can directly observe the projection-distortion in the post-specimen lenses of probe-corrected instruments, the 3-fold caustic when an image-corrector is fitted, and any misalignment of imaging detectors or spectrometer apertures. We also discuss for the first time, the effect that higher-order aberrations in the objective-lens pre-field has on such an angle-based detector mapping procedure.	Jan 2018
	Biological applications of cryo-soft X-ray tomography E. Duke , K. Dent , M. Razi , L. M. Collinson Journal Of Microscopy 255 (2), 65-70 DOI: 10.1111/jmi.12139 PMID: 24942982 Show Abstract ▼ Abstract: X-rays are used for imaging many different types of biological specimen, ranging from live organisms to the individual cells and proteins from which they are made. The level of detail achieved as a result of the imaging varies depending on both the sample and the technique used. One of the most recent technical developments in X-ray imaging is that of the soft X-ray microscope, designed to allow the internal structure of individual biological cells to be explored. With a field of view of ∼10–20 × ∼10–20 μm, a penetration depth of ∼10 μm and a resolution of ∼40 nm3, the soft X-ray microscope neatly fits between the imaging capabilities of light and electron microscopes.	Jun 2014
B16-Test Beamline	Region-of-interest tomography using filtered backprojection: assessing the practical limits Albrecht Kyrieleis , Valeriy Titarenko , Thomas Connolley , Mark Ibison , Philip Withers Journal Of Microscopy DOI: 10.1111/j.1365-2818.2010.03408.x PMID: 21118206 Diamond Proposal Number(s): [486] Show Abstract ▼ Abstract: There are many cases where it is desirable to reconstruct at high resolution a small volume from a larger sample. Here we describe the outcomes of a reconstruction trial based on real samples aimed at delineating the practical limits to which a small region of interest can be viewed from a large sample. Our approach has been to artificially truncate the sinograms of whole sample scans to simulate region of interest tomography. A simple filtered back projection algorithm has been applied, with the sinograms extended laterally in a simple manner to make up for the truncated portions. The impact of the degree of truncation (from 0% down to 99%), the number of projections used, as well as the position of the region of interest, on the faithfulness of the reconstruction is evaluated for a range of sample types. We have assessed the nature of, and extent to which, artefacts are introduced and the degree to which simple strategies can minimize them to an acceptable level without the need for complex reconstruction algorithms, projection stitching strategies or very large detectors. It is found that for a wide range of objects the effect of truncation on feature detection is negligible and that excellent images can be reconstructed if the number of projections is calculated not on the basis of the number of pixels on the camera, but on the number of pixels that would be required to scan the whole sample at the chosen pixel resolution. This paper demonstrates that in many cases more sophisticated reconstruction strategies are not necessary.	Jun 2010

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