Bronchoalveolar Lavage and Lung Imaging

At killing, collection of BAL fluid was performed using 1 ml of sterile Hanks’ balanced saline buffer. The BAL protein concentration was determined by a BCATM Protein Assay kit (Thermo Scientific, Pittsburg, PA). BAL inflammatory cell counting was performed using a standard hemacytometer technique (Fu et al., 2009 (link)). As an additional parameter reflecting increased lung vascular leakiness, Evans blue accumulation in the lung tissue was evaluated as described elsewhere (Fu et al., 2009 (link)). At the end of the experiment, a thoracotomy was performed, and the lungs were perfused in situ via the left atrium with PBS containing 5 mM EDTA to flush the blood off the lungs. The left lung and right lung were excised and imaged by a Kodak digital camera.