Multiscale three-dimensional imaging of intact human organs down to the cellular scale using hierarchical phase-contrast tomography

Abstract

Human organs are complex, three-dimensional and multiscale systems. Spatially mapping the human body down through its hierarchy, from entire organs to their individual functional units and specialised cells, is a major obstacle to fully understanding health and disease. To meet this challenge, we developed hierarchical phase-contrast tomography (HiP-CT), an X-ray phase propagation technique utilising the European Synchrotron Radiation Facility's Extremely Brilliant Source: the world's first high-energy 4th generation X-ray source. HiP-CT enabled three-dimensional and non-destructive imaging at near-micron resolution in soft tissues at one hundred thousand times the voxel size whilst maintaining the organ's structure. We applied HiP-CT to image five intact human parenchymal organs: brain, lung, heart, kidney and spleen. These were hierarchically assessed with HiP-CT, providing a structural overview of the whole organ alongside detail of the organ's individual functional units and cells. The potential applications of HiP-CT were demonstrated through quantification and morphometry of glomeruli in an intact human kidney, and identification of regional changes to the architecture of the air-tissue interface and alveolar morphology in the lung of a deceased COVID-19 patient. Overall, we show that HiP-CT is a powerful tool which can provide a comprehensive picture of structural information for whole intact human organs, encompassing precise details on functional units and their constituent cells to better understand human health and disease.

Progress on In Situ and Operando X-ray Imaging of Solidification Processes

“…The synchrotron facilities and techniques are undergoing massive upgrades that will enable imaging of large components, including human body parts, at unprecedented resolutions and levels. For example, ESRF has rebuilt the brightest-to-date X-ray source, ([ 166 ]), and the same facility has demonstrated new hierarchical phase-contrast imaging techniques ([ 167 ]). Such advances will enable scanning of a full aero or automobile engine or replicate large-scale phenomena in an imaging beamline.…”

3D virtual histopathology of cardiac tissue from Covid-19 patients based on phase-contrast X-ray tomography

“…with sub-micron voxel size and with sufficient contrast also for soft and unstained tissues (Töpperwien et al, 2018). By relaxing the resolution to voxel sizes in the range of 25 microns and stitching of different tomograms, the entire human organ can be covered and an entire FFPE tissue block 'unlocked' by destruction-free 3d analysis (Walsh et al, 2021) In this work, we now focus on the 3d architecture of cardiac tissue. We have scanned unstained, paraffin embedded tissue, prepared by a biopsy punch from paraffin embedded tissue blocks, collected from patients which have succumbed to .…”

3D virtual Histopathology of Cardiac Tissue from Covid-19 Patients based on Phase-Contrast X-ray Tomography

“…More generally, one would like to delineate the morphological changes of cytoarchitecture from other well described pathologies. Recently, we have introduced three-dimensional (3d) virtual histology based on phase-contrast x-ray tomography as a new tool for Covid-19 pathohistology and investigated these structural changes in post mortem tissue biopsies from Covid-19 diseased lung tissue using propagation based x-ray tomography (Eckermann et al, 2020;Walsh et al, 2021). Exploiting phase contrast based on wave propagation, the 3d structure of formalin-fixed, paraffin-embedded (FFPE) tissue -the mainstay for histopathological samples worldwide-can be assessed at high resolution, i.e.…”

“…with sub-micron voxel size and with sufficient contrast also for soft and unstained tissues (Töpperwien et al, 2018). By relaxing the resolution to voxel sizes in the range of 25 microns and stitching of different tomograms, the entire organ can be covered and an entire FFPE tissue block 'unlocked' by destruction-free 3d analysis (Walsh et al, 2021) In this work, we now focus on the 3d cytoarchitecture of cardiac tissue. We have scanned unstained, paraffin embedded tissue, prepared by a biopsy punch from paraffin embedded tissue blocks, collected from patients which have succumbed to .…”