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Optimising complementary soft tissue synchrotron X-ray microtomography for reversibly-stained central nervous system samples

DOI: 10.1038/s41598-018-30520-8 DOI Help

Authors: Merrick C. Strotton (King's College London) , Andrew J. Bodey (Diamond Light Source) , ( Joint First Authors ) , Kazimir Wanelik (Diamond Light Source) , Michele C. Darrow (Diamond Light Source) , Esau Medina (King’s College London) , Carl Hobbs (King’s College London) , Christoph Rau (Diamond Light Source) , Elizabeth J. Bradbury (King’s College London)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Scientific Reports , VOL 8

State: Published (Approved)
Published: August 2018
Diamond Proposal Number(s): 12538 , 14907

Open Access Open Access

Abstract: Synchrotron radiation microtomography (SRμCT) is a nominally non-destructive 3D imaging technique which can visualise the internal structures of whole soft tissues. As a multi-stage technique, the cumulative benefits of optimising sample preparation, scanning parameters and signal processing can improve SRμCT imaging efficiency, image quality, accuracy and ultimately, data utility. By evaluating different sample preparations (embedding media, tissue stains), imaging (projection number, propagation distance) and reconstruction (artefact correction, phase retrieval) parameters, a novel methodology (combining reversible iodine stain, wax embedding and inline phase contrast) was optimised for fast (~12 minutes), high-resolution (3.2–4.8 μm diameter capillaries resolved) imaging of the full diameter of a 3.5 mm length of rat spinal cord. White-grey matter macro-features and micro-features such as motoneurons and capillary-level vasculature could then be completely segmented from the imaged volume for analysis through the shallow machine learning SuRVoS Workbench. Imaged spinal cord tissue was preserved for subsequent histology, establishing a complementary SRμCT methodology that can be applied to study spinal cord pathologies or other nervous system tissues such as ganglia, nerves and brain. Further, our ‘single-scan iterative downsampling’ approach and side-by-side comparisons of mounting options, sample stains and phase contrast parameters should inform efficient, effective future soft tissue SRμCT experiment design.

Subject Areas: Technique Development, Biology and Bio-materials, Physics

Diamond Offline Facilities: I13 Data Beamline
Instruments: I13-2-Diamond Manchester Imaging