Methods in Pulmonary Research

Airways.- 1 Measurement of lung function in rodents in vivo.- Spontaneous respiration.- Pulmonary manoeuvres.- Material and equipment.- Lung function laboratory.- Methods.- Preparation and calibration.- Pulmonary function testing.- Examples for applications.- Discussion.- Troubleshooting.- References.- 2 The isolated perfused lung.- Advantages and disadvantages of perfused lungs.- Theoretical background.- Vascular resistance.- Respiratory mechanics.- Material and equipment.- Artificial thorax chamber and ventilation.- Perfusion.- Weight measurement.- Gas exchange.- Methods.- Surgery and setting up the lung.- Criteria for viability.- Cleaning the apparatus.- An example application.- Discussion.- Interpretation of the results.- Constant flow (CFP) versus constant pressure perfusion (CPP).- Negative or positive pressure ventilation.- Choice of perfusate.- Recirculating versus non-recirculating perfusion.- Additional experimental options.- Troubleshooting.- Final comments.- References.- 3 Lung explants.- Material and equipment.- Preparation of culture media.- Preparation of agarose.- Preparation of animals.- Preparation of explants.- Image acquisition.- Variations on this technique.- Applications.- Effects of bronchoconstriction.- Measurements of mucociliary clearance.- Measurements of pulmonary vasculature.- Long term explant culture techniques.- Investigations of protein and gene expression.- Troubleshooting.- Discussion.- Acknowledgements.- References.- 4 Tracheal preparations.- Methods.- Guinea pig tracheal preparations.- Immersion techniques.- Tracheal chain.- Spirally cut trachea.- Zig-zag tracheal strip.- Tracheal tube preparations.- Superfusion techniques.- Electrically stimulated trachea.- Epithelium-denuded trachea.- Conclusion.- References.- Vessels.- 5 Intravital microscopy: Airway circulation.- Materials and equipment.- Microscope.- Video equipment.- Peripheral equipment.- Ventilation.- Solutions.- Methods.- Surgery.- Experimental procedure.- Species differences.- Discussion.- References.- 6 The bronchial circulation.- Importance and role of the bronchial circulation.- Postobstructive pulmonary vasculopathy (POPV) and principles of the techniques.- Material and equipment.- Production of POPV in dogs, rats and guinea pigs: Ligation of the left main pulmonary artery.- In situ perfused LLL preparation.- Morphological assessment of the bronchial and pulmonary vasculature using light microscopy and morphometry.- Methods.- Surgical ligation of the left main pulmonary artery in dogs, rats and guinea pigs.- Canine model.- Rat and guinea pig model.- In situ perfused LLL preparation to measure pulmonary and bronchial vascular flows, pressures and resistances using modified AO and VO and bronchial vascular micropuncture.- Procedure for the in situ perfused LLL preparation.- AO and VO measurements.- Modified in situ perfused LLL preparation for bronchial collateral.- vascular pressure measurements by micropuncture.- Morphological assessment of the bronchial and pulmonary vasculature, using light microscopy and morphometry.- Measurement of pulmonary vascular medial thickness and muscularization in lungs injected with pigmented gelatin-barium mixtures.- Fixation and preparation.- Morphometry.- Assessment of proliferation in the bronchial vasculature.- Bronchial vessel number per airway.- Assessment of bronchial vascular endothelial proliferation using bromodeoxyuridine (BrdU) labeling.- Discussion and troubleshooting.- Production of POPV.- In situ perfused left lower lobar preparation.- Morphological assessment of the bronchial and pulmonary vasculature.- Acknowledgements.- References.- 7 Segmental vascular resistance and compliance from vascular occlusion.- Methods.- The lumped parameter RCR model.- The continuous RC distribution.- More distributed lumped parameter models.- The 3C4R model.- The 3C2R model.- Arterial occlusion in vivo.- Acknowledgements.- References.- Edema.- 8 Experimental and clinical measurement of pulmonary ...