Abstract
In this study we have utilized an optical clearing method to allow visualization of a heretofore undescribed subpleural acinar structural organization in the mammalian lung. The clearing method enables visualization of the lung structure deep below the visceral pleura in intact inflated lungs. In addition to confirming previous observations that the immediate subpleural alveoli are uniform in appearance, we document for the first time that the subpleural lung parenchyma is much more uniformly organized than the internal parenchyma. Specifically, we report that below the surface layer of alveoli, there is a striking parallel arrangement of alveolar ducts that all run perpendicular to the visceral pleural surface. A three dimensional visualization of alveolar ducts allowed for a calculation of the average inner to outer duct diameter ratio of 0.53 in these subpleural ducts. This unique, self-organizing parallel duct structure likely impacts both elastic recoil and the transmission of tethering forces in healthy and diseased lungs.
Highlights
In this study we have utilized an optical clearing method to allow visualization of a heretofore undescribed subpleural acinar structural organization in the mammalian lung
Our studies were initially focused on mouse lungs, but since the distinctive organization we found had not been described, we confirmed in other species that this pattern was not unique to the mouse
We found for the uncleared tissue that, while the initial surface layer of alveoli appeared similar to that of the cleared lungs, with confocal imaging beyond a depth of about ≈60 μm it was impossible to clearly define any of the lung structures
Summary
In this study we have utilized an optical clearing method to allow visualization of a heretofore undescribed subpleural acinar structural organization in the mammalian lung. A three dimensional visualization of alveolar ducts allowed for a calculation of the average inner to outer duct diameter ratio of 0.53 in these subpleural ducts This unique, selforganizing parallel duct structure likely impacts both elastic recoil and the transmission of tethering forces in healthy and diseased lungs. It is clear from light microcopy that there are alveoli immediately under the pleural surface, there has been no information of how the alveolar duct might be oriented in this region. The optical clearing method permits imaging in the mouse lung to depths exceeding 30 alveolar diameters into the lung, allowing optical serial sectioning of the subpleural alveoli and the ducts beneath them. Results in all species show that all of the most peripheral alveolar ducts are oriented perpendicular to the visceral pleural surface
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