Introduction Intravital microscopy (IVM) is the gold standard for in vivo investigations of the microcirculation. The availability of transgenic animals has propelled the development of murine IVM models for various organs, but not yet for the lung. We report a new IVM model for studies of the murine pulmonary microcirculation. Method Balb/c mice were anesthetized, tracheotomized, and ventilated mechanically. Catheters were placed in the right jugular vein and carotid artery for hemodynamic monitoring. A window was surgically excised from the right thoracic wall exposing the upper lobe of the right lung. The window was closed with a transparent polyvinylidene membrane and sealed with α-cyanoacrylate. Intrathoracic air was removed via a transabdominal intrapleural catheter. FITC-dextran perfused microvessels, rhodamine 6G-stained leukocytes, or platelets labeled ex vivo by CFDA were imaged. Results IVM preparations were hemodynamically stable with mean arterial pressures > 70 mmHg, arterial blood gases were in the physiological range with PO2 of 91±4 mmHg and PCO2 of 34±2 mmHg after 120 min (n=5). Wet/dry ratio of imaged lungs (4.4±0.2, n=4) did not differ from control lungs (4.4±0.1, n=3). Within each window of observation, 4–5 arterioles and 2–3 venules with diameters ranging between 56-21 μm (arterioles) and 34-16 μm (venules) could be stably visualized. Circulating and adherent blood cells could be identified and quantified in the different segments of the pulmonary microvasculature. Conclusion This model provides a stable murine pulmonary IVM preparation under closed thorax conditions with no signs of impaired gas exchange or edema formation. It presents a powerful new technique for the study of pulmonary microvascular physiology and pathology in transgenic mice. Supported by Tyojyu Kagaku and Kaiserin Friedrich Foundation
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