Real-time imaging of cellular and sub-cellular dynamics in vascularized organs requires image-resolution, image-registration, and demonstrably intact physiology to be simultaneously optimized. This problem is particularly pronounced in the lung in which cells may transit at speeds > 1 mm/sec, and in which normal respiration results in large-scale tissue movements that prevent image registration. Here, we report video-rate, two-photon imaging of a physiologically intact preparation of the mouse lung that is at once stabilizing and non-disruptive. The application of our method provides evidence for differential trapping of T cells and neutrophils in mouse pulmonary capillaries and enables observation of neutrophil mobilization and dynamic vascular leak in response to stretch and inflammatory models of lung injury in mice. The system permits physiological measurement of motility rates of > 1 mm/sec, observation of detailed cellular morphology, and could be applied to other organs and tissues while maintaining intact physiology.

This review provides an overview of the relationship between ventilation/perfusion ratios and gas exchange in the lung, emphasising basic concepts and relating them to clinical scenarios. For each gas exchanging unit, the alveolar and effluent blood partial pressures of oxygen and carbon dioxide (PO 2 and PCO 2 ) are determined by the ratio of alveolar ventilation to blood flow (V9A/Q9) for each unit. Shunt and low V9A/Q9 regions are two examples of V9A/Q9 mismatch and are the most frequent causes of hypoxaemia. Diffusion limitation, hypoventilation and low inspired PO 2 cause hypoxaemia, even in the absence of V9A/Q9 mismatch. In contrast to other causes, hypoxaemia due to shunt responds poorly to supplemental oxygen. Gas exchanging units with little or no blood flow (high V9A/Q9 regions) result in alveolar dead space and increased wasted ventilation, i.e. less efficient carbon dioxide removal. Because of the respiratory drive to maintain a normal arterial PCO 2 , the most frequent result of wasted ventilation is increased minute ventilation and work of breathing, not hypercapnia. Calculations of alveolar-arterial oxygen tension difference, venous admixture and wasted ventilation provide quantitative estimates of the effect of V9A/Q9 mismatch on gas exchange. The types of V9A/Q9 mismatch causing impaired gas exchange vary characteristically with different lung diseases. @ERSpublications A review of ventilation-perfusion relationships and gas exchange, basic concepts and their relation to clinical cases

Estimations of dog lung, pig heart, and pig kidney regional perfusion by use of fluorescent-labeled microspheres were compared with measurements obtained with standard radiolabeled microspheres. Pairs of radio- and fluorescent-labeled microspheres (15 microns diam, 6 colors) were injected into a central vein of a supine anesthetized dog and the left ventricle of three supine anesthetized pigs while reference blood samples were simultaneously withdrawn from a femoral artery in the pigs. The lungs were cubed into approximately 2 cm3 pieces (n = 1,510). Each pig heart and kidney was cubed into approximately 1-g pieces (total n = 192 and 120, respectively). The radioactivity of each organ piece and reference blood sample was determined using a scintillation counter with count rates corrected for decay, background, and spillover. Tissue samples and reference blood samples were digested with KOH and filtered and the fluorescent dye was extracted with a solvent, or the dye was extracted from lung tissue without filtering. The fluorescence of each sample was determined for each color by use of an automated spectrophotometer. Perfusion was calculated for each organ piece from both the radioactivity and fluorescence. Correlation between flow determined by radio- and fluorescent-labeled microspheres was as follows: r = 0.96 +/- 0.01 (SD) (lung, filtered, n = 588), r = 0.99 +/- 0.00 (lung, nonfiltered, n = 710), r = 0.95 +/- 0.02 (heart, filtered), and r = 0.96 +/- 0.02 (kidney, filtered). Compared with colored microspheres, methods for quantitating fluorescent-labeled microspheres are more sensitive, less labor intensive, and less expensive. Fluorescent-labeled microspheres provide a new nonradioactive method for single and repeated measurement of regional organ perfusion.

No abstract