Hard X-ray submicrometer tomography of human brain tissue at Diamond Light Source

X-Ray Microscopy Conference 2016

Khimchenko, A.^1 ; Bikis, C.^1 ; Schulz, G.^1 ; Zdora, M.-C.^2,3 ; Zanette, I.^2 ; Vila-Comamala, J.^2 ; Schweighauser, G.^4 ; Hench, J.^4 ; Hieber, S.E.^1 ; Deyhle, H.^1 ; Thalmann, P.^1 ; Müller, B.^1

Biomaterials Science Center, Department of Biomedical Engineering, University of Basel, Allschwil, Switzerland^1

Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire, Didcot

OX11 0DE, United Kingdom^2

Department of Physics and Astronomy, University College London, London

WC1E 6BT, United Kingdom^3

Institute of Pathology, Department of Neuropathology, Basel University Hospital, Basel, Switzerland^4

关键词: Diamond light source;    High spatial resolution;    Histological section;    In-line phase contrast;    Phase reconstruction;    Subcellular structure;    Threedimensional (3-d);    Tissue transformation;   

Others  :  https://iopscience.iop.org/article/10.1088/1742-6596/849/1/012030/pdf
DOI  :  10.1088/1742-6596/849/1/012030

来源: IOP

【 摘 要 】

There is a lack of the necessary methodology for three-dimensional (3D) investigation of soft tissues with cellular resolution without staining or tissue transformation. Synchrotron radiation based hard X-ray in-line phase contrast tomography using single-distance phase reconstruction (SDPR) provides high spatial resolution and density contrast for the visualization of individual cells using a standard specimen preparation and data reconstruction. In this study, we demonstrate the 3D characterization of a formalin-fixed paraffin-embedded (FFPE) human cerebellum specimen by SDPR at the Diamond-Manchester Imaging Branchline I13-2 (Diamond Light Source, UK) at pixel sizes down to 0.45 μm. The approach enables visualization of cerebellar layers (Stratum moleculare and Stratum granulosum), the 3D characterization of individual cells (Purkinje, stellate and granule cells) and can even resolve some subcellular structures (nucleus and nucleolus of Purkinje cells). The tomographic results are qualitatively compared to hematoxylin and eosin (H&E) stained histological sections. We demonstrate the potential benefits of hard X-ray microtomography for the investigations of biological tissues in comparison to conventional histology.