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Background: A comprehensive understanding of vascular development in the human lung is still missing., Methods: Therefore, samples of infant (n = 5, 26 days to 18 months postnatally) and adult (n = 5, 20 to 40 years) human lungs were subjected to unbiased stereological estimation of the total number of capillary loops. Serial sections were segmented to visualize the alveolar capillary network (ACN) in 3D., Results: The number of capillary loops increased in parallel to lung volume from 26 days to 18 months, while in adults, it was not correlated to lung volume. In infant lungs, two capillary layers were separated by a connective tissue sheet with a growing number of interconnections. In adults, the mature ACN was almost, but not completely, single-layered. Here, the connective tissue was thinner but still centrally positioned, suggesting the persistence of interconnected parts of both layers of the previously double-layered ACN., Conclusions: Small parts of the capillaries remain double-layered and seem to be grouped around the thin connective tissue sheet, suggesting a different mechanism of microvascular maturation than simple fusion of the two layers. These spots are a potential basis for further alveolarization after completion of bulk formation., Impact: The 3D data offer a new conceptual approach to microvascular maturation of the lung. Microvascular maturation rather results from reduction than simple fusion of capillary fragments. Adult lungs maintain small double-layered capillary spots. These could offer a potential source of regeneration. The data are important to better understand normal and pathological lung development., (© 2024. The Author(s).)

Stereology, the gold standard of lung morphometry, critically depends on sampling of tissue for analysis. Random sampling approaches guarantee each part of the organ an equal chance of being included in the analysis, hence they guarantee a representative sample of the whole. However, when biological or pathological structures of interest are rare and/or heterogeneously distributed over the whole lung, the random sampling approach can be inefficient or even result in meaningless data. In such cases, a targeted sampling approach can be useful which helps to relate the analytical items to an appropriate reference space. Targeted stereology greatly benefits from the increasing availability of multi-resolution imaging techniques at macroscopic and microscopic level as well as digital tools of segmentation. As such, the present article outlines two basic sampling scenarios: 1. In the first scenario, computed tomography and microscopy are subsequently used to segment the airway/arterial tree and perform stereological measurements on specific branches of the tree. 2. The second scenario deals with heterogeneous distribution of pathological lesions. This type of analysis can be divided into two stages: assessment of lesions of interest (LOI) within the lung and assessment of subcompartments within LOI. Taken together, targeted stereology has a thorough foundation in stereological theory and is not only able to significantly increase the efficiency of the analysis but also to yield new types of information that would be lost with the classical random sampling approach.

Exocrine and endocrine pancreas are interconnected anatomically and functionally, with vasculature facilitating bidirectional communication. Our understanding of this network remains limited, largely due to two-dimensional histology and missing combination with three-dimensional imaging. In this study, a multiscale 3D-imaging process was used to analyze a porcine pancreas. Clinical computed tomography, digital volume tomography, micro-computed tomography and Synchrotron-based propagation-based imaging were applied consecutively. Fields of view correlated inversely with attainable resolution from a whole organism level down to capillary structures with a voxel edge length of 2.0 µm. Segmented vascular networks from 3D-imaging data were correlated with tissue sections stained by immunohistochemistry and revealed highly vascularized regions to be intra-islet capillaries of islets of Langerhans. Generated 3D-datasets allowed for three-dimensional qualitative and quantitative organ and vessel structure analysis. Beyond this study, the method shows potential for application across a wide range of patho-morphology analyses and might possibly provide microstructural blueprints for biotissue engineering., (© 2024. The Author(s).)