Interchangeable lens digital camera

BAL fluid samples were collected by wedging a 3.8 mm flexible bronchoscope (aScope 3, Ambu) into a subsegmental bronchus of an injured or control lung. Sterile normal saline (5 mL) was injected, aspirated, and collected in a sterile specimen trap (Busse Hospital Disposables). Gross images of BAL fluid samples were acquired throughout 36 h of cross-circulation using an interchangeable lens digital camera (Sony). The pH of BAL fluid samples was measured with a glass double junction microelectrode (Fisher Scientific). BAL fluid samples were centrifuged at 2164 × g for 10 min at 4 °C. Snap-frozen supernatants were stored at –80 °C until further processing.

Following unilateral bronchoscopic delivery of standardized gastric contents into donor lungs, donor vitals and ventilator settings were continuously recorded. Chest X-rays, airway bronchoscopic images, and hemogases were obtained at baseline and every 2 h for 6 h. Donor blood samples were collected at baseline and 6 h after gastric aspiration to assess serum levels of inflammatory markers (GM-CSF, IFNγ, IL-1α, IL-1β, IL-1rα, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18, P-Selectin, and TNFα) by multiplex array analysis (Eve Technologies) and hemolytic markers (D-dimer, fibrinogen, and plasma free hemoglobin) by commercially available enzyme-linked immunosorbant assays (ELISAs). A list of the ELISA kits used is provided in Supplementary Table 11. BAL fluid samples were collected from injured and control lungs at baseline and 6 h after gastric aspiration to assess the airway response to gastric aspiration. Inflammatory markers (GM-CSF, IFNγ, IL-1α, IL-1β, IL-1rα, IL-2, IL-4 IL-6, IL-8, IL-10, IL-12, IL-18, and TNFα) in BAL fluid samples were quantified by multiplex array analysis (Eve Technologies), M30 by commercially available ELISA, and cellular contents in BAL fluid were analyzed by Kwik-Diff staining or periodic acid-Schiff staining. Gross images of injured lungs were obtained prior to explant using an interchangeable lens digital camera (Sony).