To supply the large amount of capillaries, the pulmonary arteries form a tree-like structure with numerous branches of decreasing diameter. The mammalian pulmonary vasculature consists of an arterial inflow, a vast capillary bed and one or more veins draining the blood from the lungs. In conclusion, the correlative image analysis of ♜T and LM-based datasets is an innovative method to assess the pulmonary vasculature quantitatively. Stereological analysis for the first three arterial generations of the monopodial branching of the vasculature included volume fraction, total volume, lumen-to-wall ratio and wall thickness for each arterial generation. The present study demonstrates that it is feasible to identify arterial profiles according to their generation based on a generation-specific color code. The digital processing involved segmentation of the arterial tree, generation analysis, registration of LM sections with the ♜T data as well as registration of the segmentation and the LM images. The lung was then exhaustively sectioned and 20 consecutive sections were collected every 100 µm to obtain a systematic uniform random sample of the whole lung. A whole left rabbit lung was fixed by vascular perfusion, embedded in glycol methacrylate and imaged by micro-computed tomography (♜T). Therefore, we established a workflow that allows for the generation-specific quantitative (stereological) analysis of pulmonary blood vessels. The light microscopic (LM) analysis of the blood vessels is limited by the fact that it is impossible to identify which generation of the arterial tree an arterial profile within a LM microscopic section belongs to. Various lung diseases, including pulmonary hypertension, chronic obstructive pulmonary disease or bronchopulmonary dysplasia, are associated with structural and architectural alterations of the pulmonary vasculature.
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