One approach is optical clearing, which homogenizes the refractive index in biological tissues, allowing high-resolution 3D imaging modalities such as light-sheet microscopy or optical projection tomography. In recent years, there has been progress toward achieving 3D imaging of intact organs at multiple length scales. New 3D imaging techniques are therefore required to bridge length scales from spatial relationships at the cellular level to the architectural organization of intact human organs. Physical subsampling creates challenges in data registration and the requirement that correct or representative subsamples are collected 2, 3. Currently, hierarchical imaging typically involves physical subsampling of larger samples before high-resolution imaging 2. A more practical approach is to provide overall spatial context at a lower resolution and then use this data to select smaller regions for higher-resolution imaging this type of imaging can be considered hierarchical imaging. Spatial mapping at the single-cell level for entire human organs is presently unfeasible in terms of existing techniques, data-storage requirements and analysis or interpretability 1. Thus, mapping the spatial organization and morphology of individual cells up to the scale of intact organs is fundamental to understanding system-level behaviors in health or disease. Spatial relationships, 3D morphology and interaction within and across these length scales collectively provide a basis for biological function. We demonstrate the potential applications of HiP-CT through quantification and morphometry of glomeruli in an intact human kidney and identification of regional changes in the tissue architecture in a lung from a deceased donor with coronavirus disease 2019 (COVID-19).īiological tissues are complex 3D structures arranged hierarchically from individual specialized cells to organotypic functional units, for example, alveoli in the lung up to intact, whole organs. HiP-CT provided a structural overview of each whole organ followed by multiple higher-resolution volumes of interest, capturing organotypic functional units and certain individual specialized cells within intact human organs. We applied HiP-CT to image five intact human organ types: brain, lung, heart, kidney and spleen. The spatial coherence of the ESRF-EBS combined with our beamline equipment, sample preparation and scanning developments enabled us to perform non-destructive, three-dimensional (3D) scans with hierarchically increasing resolution at any location in whole human organs. ![]() To meet this challenge, we developed hierarchical phase-contrast tomography (HiP-CT), an X-ray phase propagation technique using the European Synchrotron Radiation Facility (ESRF)’s Extremely Brilliant Source (EBS). Imaging intact human organs from the organ to the cellular scale in three dimensions is a goal of biomedical imaging.
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