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Bellier, A.

Publications and source records attributed to Bellier, A..

18 recordsLinked to original sources

Revealing multiscale architecture of the intact human spine using hierarchical phase contrast tomography

Background: Computed tomography (CT) and magnetic resonance imaging (MRI) provide detailed clinical assessment of the human spine; however, their millimeter-scale spatial resolution limits direct visualization of tissue microarchitecture and interfaces. Hierarchical phase-contrast tomography (HiP-CT) enables ex vivo non-destructive, micron-resolution imaging of intact human organs without exogenous contrast agents, extending anatomical assessment to spatial scales inaccessible to conventional clinical imaging. Purpose: To evaluate the capability of HiP-CT to visualize macro- to microanatomy of an intact adult human spine ex vivo. Methods: An intact spine (C1-L5) from a 62-year-old male donor without known skeletal disease was formalin-fixed and imaged by HiP-CT at the European Synchrotron. Whole-spine imaging was performed at 16.26 m isotropic voxels, with selected regions of interest imaged using 2.2 m voxels. Ex vivo MRI using clinical sequences was performed for anatomical comparison. Three-dimensional segmentation and quantitative analyses included intervertebral disc (IVD) grading, annulus fibrosus fiber tracing, and vertebral trabecular micro-morphometry. Results: HiP-CT enabled high spatial-resolution, multiscale imaging of the intact human spine, simultaneously visualizing vertebral bone and endplate, IVDs, spinal cord and meninges, nerve roots and ganglia, vasculature, ligaments, and musculature. Three-dimensional segmentation and virtual sectioning mapped neurovascular anatomy and vertebral vascular supply, without exogenous contrast enhancement, while preserving their relationships with surrounding tissues. Whole-spine images enabled quantitative analysis of annulus fibrosus fiber organization and vertebral trabecular microarchitecture, permitting the detection of focal posterolateral fiber disruption at the L4-L5 IVD and regional differences in trabecular architecture between cervical and lumbar vertebral bodies. Local tomography resolved annulus fibrosus lamellae, vertebral endplate Haversian systems, and facet joint hyaline cartilage at micron resolution. Conclusions: HiP-CT enables non-destructive, multiscale three-dimensional imaging of the intact human spine while preserving tissue relationships across spatial scales and hard-soft tissue boundaries. HiP-CT could provide an ex vivo microanatomical reference for future correlation with clinical imaging investigations of spinal disease and in the development and validation of imaging biomarkers.

bioengineering↗

BigHeart: Mapping connectivity in the adult human heart at the micron scale

The human heart depends on coordinated muscular, electrical, vascular, lymphatic, and neural systems, but their three-dimensional relationships remain unresolved at microscopic resolution across an intact adult organ. Using hierarchical phase-contrast tomography, we generated a continuous three-dimensional structural reference of a whole adult human heart at 8.01 micrometer isotropic voxel size, without sectioning or staining. Organ-wide imaging revealed hierarchical myocardial architecture, and quantitative orientation analysis identified regional differences in myocardial cell aggregate organization. The atrioventricular conduction axis was traced into a distinct subendocardial Purkinje network interfacing with working myocardium. Integrated segmentation showed that coronary vessels, lymphatic collectors, and autonomic nerves occupy shared epicardial corridors. This publicly accessible dataset integrates cardiac systems within a common spatial framework for anatomy, computational modeling, and future molecular atlases.

physiology↗

Hierarchical X-ray microscopy and mesoscopic diffusion MRI in the same brain reveal the human connectome across scales

We present a multimodal pipeline for 3D imaging of cerebral white-matter archi-tecture across scales, from whole-brain axonal projections down to individual myelinated axons. After diffusion MRI, an adult ex vivo human hemisphere undergoes label-free imaging with Hierarchical Phase-Contrast Tomography (HiP-CT) from 20 {micro}m/voxel in the whole hemisphere to 2 {micro}m/voxel in areas of interest, with intrinsic cross-scale alignment. A 4 cm tissue block extracted from the hemisphere is reimaged with HiP-CT at 0.857 {micro}m/voxel, enabling direct visualisation of single myelinated axons. After osmium staining, micro-CT at 0.364 {micro}m/voxel and electron microscopy at 4 nm/voxel are acquired in biop-sies from the tissue block to validate the presence of myelinated axons in the label-free HiP-CT contrast. Spanning three orders of magnitude in resolution, these co-registered multimodal datasets bridge microscopic wiring and macro-scopic brain organisation, providing a foundation for anatomically grounded whole-brain connectomics.

neuroscience↗

Neuroanatomy of the clitoris

The clitoris is one of the least studied organs of the human body. The detailed anatomy of the clitoris is challenging to address through a gross dissection, as most of its parts are embedded internally, surrounded by pubic bone and several pelvic organs. While clinical imaging methods such as magnetic resonance imaging can capture the gross 3D morphology, they lack the spatial resolution required to resolve the detailed structures. In this study, we generated micron-scale computed tomography images of the female pelvises, leveraging a synchrotron radiation X-ray source. This unique data revealed the complex trajectory of the dorsal nerve of the clitoris, the main sensory nerve of the clitoris. Notably, the nerve trunks within the clitoral glans were revealed, with the maximum diameter ranging from 0.2 to 0.7 mm. They showed a tree-like branching pattern projecting towards the surface of the glans. We also revealed that some branches of the dorsal nerve of the clitoris ramify to innervate the clitoral hood and mons pubis. Finally, the posterior labial nerve, a branch of the perineal nerves, was shown to innervate the surroundings of the clitoris and the labial structures. These findings have an immediate impact on operations performed around the vulva area, such as gender-affirmation surgery and reconstruction surgery after genital mutilation.

neuroscience↗

The Extremely Brilliant Brain: An Isotropic Microscale Human Brain Dataset

We present an isotropic 7.72 {micro}m/voxel post-mortem human brain dataset acquired using Hierarchical Phase-Contrast Tomography (HiP-CT) at the ESRF Extremely Brilliant Source, beamline BM18. This fills a critical gap between whole-brain MRI at 100 {micro}m resolution and serial-section histological reconstructions at 20 {micro}m or finer. HiP-CT contrast, derived from X-ray phase shifts, enables rich 3D visualisation of complex neuroanatomy including white-matter bundles, microvasculature, and sub-nuclei. We provide open-source workflows for online data exploration, subvolume download, segmentation, and reintegration of analyses into the full dataset. We demonstrate the potential of this resource by tracing vasculature over long distances, segmenting nuclei, and extracting whitematter orientations with 3D structure-tensor analysis. High-resolution human brain datasets are transformative for quantitative neuroanatomy, circuit mapping, and validation of clinical imaging; this openly available resource is a critical step for global access to next-generation multiscale brain imaging.

neuroscience↗

Hierarchical Phase-Contrast Tomography Imaging: Applicability in biomedical research

ObjectivesHierarchical Phase-Contrast Tomography (HiP-CT) enables non-destructive, multi-scale imaging of whole human organs. We describe how HiP-CT is utilized for biomedical research within the Human Organ Atlas Hub through three case studies: mapping the enteric nervous system (ENS) of the human colon, analysing myocardial and AV conduction architecture in Tetralogy of Fallot (TOF), and characterizing ductal organization in breast carcinoma. The challenges we faced with this novel biomedical data are discussed. MethodsWhole-organ and region-of-interest scans of three types of human organs were acquired at the European Synchrotron Radiation Facility (ESRF) with isotropic voxel sizes ranging from 20 {micro}m to 0.8 {micro}m. For the colon, voxel binning and RootPainter were employed to tackle data size to segment the ENS. For the heart, voxel-wise myocyte orientation mapping was calculated in terabyte-scale datasets with a high-performance computational framework (Cardiotensor). Breast carcinoma samples were correlated with histopathology for structure validation. ResultsHiP-CT revealed the large-scale organization of the ENS in the colon, enabling visualisation of the 3D structures of the ENS across the colon In TOF hearts, analysis uncovered abnormal myocardial structure and heterogeneous conduction system morphology. In breast carcinoma, HiP-CT resolved the full hierarchy of ductal structures and vascular relationships within tumour and peritumoral regions. ConclusionsHiP-CT provides unprecedented, hierarchical insight into intact human organ structure, bridging the gap between histology and radiology. Advances in knowledgeHiP-CT establishes a new ex vivo radiological modality capable of linking microscale pathology to whole-organ context, advancing translational research in neurogastroenterology, cardiology, and oncology

biophysics↗

Evolution of Hierarchical Phase-Contrast Tomography on the European Synchrotron beamlines BM05 and BM18: a whole adult human brain imaging case study

Hierarchical Phase-Contrast Tomography (HiP-CT) was recently developed to enable the ex-vivo imaging of human organs at multiple scales from whole organ down to cellular level. Using whole adult human brain imaging as a case study, this manuscript shows the evolution of this technique from its initial development at the BM05 beamline to its transition and current status at BM18. Thanks to the higher coherence, larger beam size, higher energies and larger propagation distances available at BM18 and due to the European Synchrotrons Extremely Brilliant Source upgrade (ESRF-EBS), this transition resulted in significantly improved data quality, resolution, sensitivity and speed. More recently, the implementation of a new generation of larger sCMOS cameras, helical scanning (including dedicated reconstruction algorithm developments), binning at the chip and projections levels, and the design of high-efficiency optics allowed to progressively improve the trade-off between dose and image quality, while simultaneously reducing scanning times. All these acquisition schemes present the current status of full organ imaging using HiP-CT and represent the constant efforts for the improvement of the technique towards the investigation of human organs in health, disease and aging. SynopsisThis manuscript shows the evolution of Hierarchical Phase-Contrast Tomography (HiP-CT) from its origins at the BM05 beamline to its transition to BM18 (ESRF-EBS). The novel hardware and scanning approach developments resulted in significantly improved data quality, resolution, sensitivity and speed. The case of whole adult brain imaging is presented to demonstrate the current possibilities of full organ imaging with local micron resolution using HiP-CT.

neuroscience↗

A whole-organ multi-scale in silico framework for human kidney haemodynamics informed by hierarchical phase-contrast tomography

Studying human kidney haemodynamics has been limited by the absence of complete vascular maps of the whole organ. Here we utilise previously generated multi-resolution hierarchical phase-contrast tomography (HiP-CT) coupled with a hybrid anatomically-grounded synthetic reconstruction to generate a full arterial-glomerular network of an intact human kidney comprising 1.6 million vessels and over 800,000 glomeruli. Using this anatomically comprehensive structure, we apply physics-based zero-dimensional haemodynamic modelling to quantify blood pressure, flow and simulated filtration rate across the entire organ. We show that the reconstructed human kidney vascular network exhibits order-dependent branching behaviour similar to that of rat kidneys, and that physiologically plausible pressure and flow patterns are recovered only when the full vascular network is represented. We further demonstrate how the kidney responds to macrovascular and microvascular perturbations, including stenosis of the large renal arteries and narrowing or ablation of afferent arterioles. Stenosis and arteriole narrowing exhibit threshold-type behaviour, with kidney perfusion and simulated glomerular filtration rate remaining largely preserved up to [~]50% narrowing, followed by sharp nonlinear declines beyond [~]70%. These predictions emerge in the absence of autoregulatory mechanisms, indicating that vascular geometry and resistance scaling alone contribute to kidney functional deterioration. Together, our framework provides the first organ-wide, data-driven model of human kidney haemodynamics and offers a foundation for future studies of kidney physiology and disease.

bioengineering↗

Resolving near-micron scale features within a whole sheep head using Hierarchical Phase-Contrast Tomography

BackgroundHierarchical Phase-Contrast Tomography (HiP-CT) was developed to image ex vivo intact human soft-tissue organs with local near-micron scale resolution. PurposeWe demonstrate the application of HiP-CT in combination with the recently developed Eikonal Phase Retrieval (EPR) for resolving anatomical features in large hard and soft tissue structures on a sheep head, and show the applicability for zooming to resolve anatomical features with near-micron scale resolution. Materials and MethodsWe imaged an entire skinned sheep head prepared in 70 % ethanol with HiP-CT at an isotropic voxel size of 16.5 {micro}m using the recently developed Eikonal Phase Retrieval (EPR) to reduce artefacts from bones. Local tomography zooms were taken in regions of interest: the eye (4.23 {micro}m voxels) and the coronal suture (2.20 {micro}m voxels). To compare to clinical imaging, we acquired T2-weighted MRI and CT of the sheep head and evaluated the contrast-to-noise ratio for differentiation of white and grey matter in the brain for all modalities. Further evaluations on HiP-CT images include structure tensor analysis for structural orientation in the brain as well as fibre tracking in the coronal suture. ResultsHiP-CT combined with EPR achieved high contrast for both hard and soft tissue. Comparison to MRI showed similar soft-tissue contrast, but much higher spatial resolution. Structure tensor analysis in the brain revealed the orientation of the major white matter bundles. In the eye, near-micron scale features such as retinal layers and the bundles in the optic nerve were visualized. Fiber tracking allowed analysis of the orientation of collagen fiber bundles in the coronal suture. ConclusionThis work highlights the potential of HiP-CT to image a complete sheep head, ex vivo, and hierarchically zoom without sectioning to resolve few-microns features locally, enabling comprehensive three-dimensional visualization of intricate cranial structures and their spatial interrelations. Summary statementTechnical developments in HiP-CT enable ex vivo X-ray imaging of an intact sheep head with local resolution to near-micron scale, demonstrating future viability on a human head. Key resultsO_LIA whole sheep head was imaged with synchrotron-based hierarchical phase-contrast tomography coupled with Eikonal Phase-retrieval with 16.5 {micro}m isotropic voxels. C_LIO_LIThe high contrast for both hard and soft tissue enabled differentiation of brain white and grey matter, the optic nerve and bone features within the skull. C_LIO_LILocal tomography zoom scans in an eye (4.23 {micro}m voxels) and coronal suture (2.20 {micro}m voxels) allowed visualization and analysis of near-micron sized features. C_LI

bioengineering↗

The Human Organ Atlas

We present the Human Organ Atlas (HOA), an open data repository making accessible multiscale 3D imaging of human organs. The repository also provides software tools and training resources enabling worldwide access, sharing, and analysis of these datasets, facilitating further research and the continued expansion of the HOA. The images are generated using a synchrotron imaging technique - Hierarchical Phase-Contrast Tomography (HiP-CT) that uses the ESRFs Extremely Brilliant Source, spanning whole organ imaging at around 20 m/voxel with local volumes of interest within the intact organs imaged down to [~] 1 m/voxel. This offers a comprehensive exploration of human anatomy, providing unparalleled insights into intricate structures and spatial relationships. The Human Organ Atlas offers researchers, clinicians, and educators a valuable resource for anatomical study, image analysis, medical education, and large-scale data mining.

bioinformatics↗

Multiscale Segmentation using Hierarchical Phase-contrast Tomography and Deep Learning

Biomedical systems span multiple spatial scales, encompassing tiny functional units to entire organs. Interpreting these systems through image segmentation requires the effective propagation and integration of information across different scales. However, most existing segmentation methods are optimised for single-scale imaging modalities, limiting their ability to capture and analyse small functional units throughout complete human organs. To facilitate multiscale biomedical image segmentation, we utilised Hierarchical Phase-Contrast Tomography (HiP-CT), an advanced imaging modality that can generate 3D multiscale datasets from high-resolution volumes of interest (VOIs) at ca. 1 {micro}m/voxel to whole-organ scans at ca. 20 {micro}m/voxel. Building on these hierarchical multiscale datasets, we developed a deep learning-based segmentation pipeline that is initially trained on manually annotated high-resolution HiP-CT data and then extended to lower-resolution whole-organ scans using pseudo-labels generated from high-resolution predictions and multiscale image registration. As a case study, we focused on glomeruli in human kidneys, benchmarking four 3D deep learning models for biomedical image segmentation on a manually annotated high-resolution dataset extracted from VOIs, at 2.58 to ca. 5 {micro}m/voxel, of four human kidneys. Among them, nnUNet demonstrated the best performance, achieving an average test Dice score of 0.906, and was subsequently used as the baseline model for multiscale segmentation in the pipeline. Applying this pipeline to two low-resolution full-organ data at ca. 25 {micro}m/voxel, the model identified 1,019,890 and 231,179 glomeruli in a 62-year-old donor without kidney diseases and a 94-year-old hypertensive donor, enabling comprehensive morphological analyses, including cortical spatial statistics and glomerular distributions, which aligned well with previous anatomical studies. Our results highlight the effectiveness of the proposed pipeline for segmenting small functional units in multiscale bioimaging datasets and suggest its broader applicability to other organ systems.

bioinformatics↗

Vasculature segmentation in 3D hierarchical phase-contrast tomography images of human kidneys

Efficient algorithms are needed to segment vasculature in new three-dimensional (3D) medical imaging datasets at scale for a wide range of research and clinical applications. Manual segmentation of vessels in images is time-consuming and expensive. Computational approaches are more scalable but have limitations in accuracy. We organized a global machine learning competition, engaging 1,401 participants, to help develop new deep learning methods for 3D blood vessel segmentation. This paper presents a detailed analysis of the top-performing solutions using manually curated 3D Hierarchical Phase-Contrast Tomography datasets of the human kidney, focusing on the segmentation accuracy and morphological analysis, thereby establishing a benchmark for future studies in blood vessel segmentation within phase-contrast tomography imaging.

bioinformatics↗

Micro to macro scale anatomical analysis of the human hippocampal arteries with synchrotron Hierarchical Phase-Contrast Tomography

PurposeTo date, no non-invasive imaging modality has been employed to profile the structural intricacies of the hippocampal arterial microvasculature in humans. We hypothesised that synchrotron-based imaging of the human hippocampus would enable precise characterisation of the arterial microvasculature. MethodsTwo preserved human brains from, a 69-year-old female and a 63-year-old male body donors were imaged using hierarchical phase-contrast tomography (HiP-CT) with synchrotron radiation at multiple voxel resolutions from 25.08 m down to 2.45 m. Subsequent manual and semi-automatic artery segmentation were performed followed by morphometric analyses. These data were compared to published data from alternative methodologies. ResultsHiP-CT made it possible to segment in context the arterial architecture of the human hippocampus. Our analysis identified anterior, medial and posterior hippocampal arteries arising from the P2 segment of the posterior cerebral artery on the image slices. We mapped arterial branches with external diameters greater than 50 m in the hippocampal region. We visualised vascular asymmetry and quantified arterial structures with diameters as small as 7 m. ConclusionsThrough the application of HiP-CT, we have provided the first imaging visualisation and quantification of the arterial system of the human hippocampus at high resolution in the context of whole brain imaging. Our results bridge the gap between anatomical and histological scales.

neuroscience↗

Multidimensional Analysis of the Adult Human Heart in Health and Disease using Hierarchical Phase-Contrast Tomography (HiP-CT)

Cardiovascular diseases (CVDs) are a leading cause of death worldwide. Current clinical imaging modalities provide resolution adequate for diagnosis but are unable to provide detail of structural changes in the heart, across length-scales, necessary for understanding underlying pathophysiology of disease. Hierarchical Phase-Contrast Tomography (HiP-CT), using new (4th) generation synchrotron sources, potentially overcomes this limitation, allowing micron resolution imaging of intact adult organs with unprecedented detail. In this proof of principle study (n=2), we show the utility of HiP-CT to image whole adult human hearts ex-vivo: one control without known cardiac disease and one with multiple known cardiopulmonary pathologies. The resulting multiscale imaging was able to demonstrate exemplars of anatomy in each cardiac segment along with novel findings in the cardiac conduction system, from gross (20 um/voxel) to cellular scale (2.2 um/voxel), non-destructively, thereby bridging the gap between macroscopic and microscopic investigations. We propose that the technique represents a significant step in virtual autopsy methods for studying structural heart disease, facilitating research into abnormalities across scales and age-groups. It opens up possibilities for understanding and treating disease; and provides a cardiac blueprint with potential for in-silico simulation, device design, virtual surgical training, and bioengineered heart in the future.

pathology↗

Micro to macro scale analysis of the intact human renal arterial tree with Synchrotron Tomography

The architecture of the kidney vasculature is essential for its function. Although structural profiling of the intact rodent kidney vasculature has been performed, it is challenging to map vascular architecture of larger human organs. We hypothesised that hierarchical phase-contrast tomography (HiP-CT) would enable quantitative analysis of the entire human kidney vasculature. Combining label-free HiP-CT imaging of an intact kidney from a 63-year-old male with topology network analysis, we quantitated vasculature architecture in the human kidney down to the scale of arterioles. Although human and rat kidney vascular topologies are comparable, vascular radius decreases at a significantly faster rate in humans as vessels branch from artery towards the cortex. At branching points of large vessels, radii are theoretically optimised to minimise flow resistance, an observation not found for smaller arterioles. Structural differences in the vasculature were found in different spatial zones of the kidney reflecting their unique functional roles. Overall, this represents the first time the entire arterial vasculature of a human kidney has been mapped providing essential inputs for computational models of kidney vascular flow and synthetic vascular architectures, with implications for understanding how the structure of individual blood vessels collectively scales to facilitate organ function.

bioengineering↗

Preparation of large biological samples for high-resolution, hierarchical, multi-modal imaging

Imaging the different scales of biological tissue is essential for understanding healthy organ behavior and pathophysiological changes. X-ray micro-tomography using both laboratory ({micro}CT) and synchrotron sources (sCT) is a promising tool to image the 3D morphology at the macro- and micro-scale of large samples, including intact human organs. Preparation of large samples for high resolution imaging techniques remains a challenge due to limitations with current methods, such as sample shrinkage, insufficient contrast, movement of the sample and bubble formation during mounting or scanning. Here, we describe a protocol to prepare, stabilize, and image large soft-tissue samples with X-ray microtomography. We demonstrate the protocol using intact human organs and Hierarchical Phase-Contrast Tomography (HiP-CT) imaging at the European Synchrotron Radiation Facility, but the protocol is equally applicable to a range of biological samples, including complete organisms, for both laboratory and synchrotron source tomography. Our protocol enhances the contrast of the sample, while preventing sample motion during the scan, even in case of different sample orientations. Bubbles trapped during mounting and those formed during scanning (in case of synchrotron X-ray imaging) are mitigated by multiple degassing steps. The sample preparation is also compatible with magnetic resonance imaging (MRI), CT, and histological observation. We describe a protocol for sample preparation and mounting which requires 25 to 34 days for a large organ such as an intact human brain or heart. This preparation time varies depending on the composition, size, and fragility of the tissue. Use of the protocol enables scanning of intact organs with a diameter of 150 mm with a local pixel size of one micron using HiP-CT.

physiology↗

A multiscale X-ray phase-contrast tomography dataset of whole human left lung

Technological advancements in X-ray imaging using bright and coherent synchrotron sources now allows to decouple sample size and resolution, while maintaining high sensitivity to the microstructure of soft, partially dehydrated tissues. The recently developed imaging technique, hierarchical phase-contrast tomography, is a comprehensive approach to address the challenge of organ-scale (up to tens of centimeters) soft tissue imaging with resolution and sensitivity down to the cellular level. Using this technique, we imaged ex vivo an entire human left lung at an isotropic voxel size of 25.08 m along with local zooms down to 6.05 - 6.5 m and 2.45 - 2.5 m in voxel size. The high tissue contrast offered by the fourth-generation synchrotron source at the European Synchrotron Radiation Facility reveals complex multiscale anatomical constitution of the human lung from the macroscopic (centimeter) down to the microscopic (micrometer) scale. The dataset provides complete organ-scale 3D information of the secondary pulmonary lobules and delineates the microstructure of lung nodules with unprecedented detail.

physiology↗

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

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 Facilitys Extremely Brilliant Source: the worlds 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 organs 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 organs 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.

physiology↗