National Jewish Health View Institution's Website 4 articles published in JoVE Neuroscience Air-Inflation of Murine Lungs with Vascular Perfusion-Fixation Stacey M. Thomas1, Joseph Bednarek2, William J. Janssen1, Patrick S. Hume1 1Division of Pulmonary, Sleep and Critical Care Medicine, Department of Medicine, National Jewish Health, 2 Presented is a method for air-inflation with vascular perfusion-fixation of the lungs that preserves the location of cells within airways, alveoli and interstitium for structure-function analyses. Constant airway pressure is maintained with an air-inflation chamber while fixative is perfused via the right ventricle. Lungs are processed for histologic studies. Immunology and Infection Detection and Enrichment of Rare Antigen-specific B Cells for Analysis of Phenotype and Function Mia J. Smith1,2, Thomas A. Packard1, Shannon K. O'Neill1, Rochelle M. Hinman1, Marynette Rihanek3, Peter A. Gottlieb3, John C. Cambier1,4 1Department of Immunology and Microbiology, University of Colorado School of Medicine, 2Department of Microbiology, Immunology, and Pathology, Colorado State University, 3Barbara Davis Center for Childhood Diabetes, University of Colorado School of Medicine, 4Department of Biomedical Research, National Jewish Health A simple yet effective method that employs magnetic nanoparticles to detect and enrich antigen-reactive B cells for functional and phenotypic analysis is described. Bioengineering Multi-step Preparation Technique to Recover Multiple Metabolite Compound Classes for In-depth and Informative Metabolomic Analysis Charmion Cruickshank-Quinn1, Kevin D. Quinn1, Roger Powell1, Yanhui Yang1, Michael Armstrong1, Spencer Mahaffey2, Richard Reisdorph1, Nichole Reisdorph1 1Department of Immunology, National Jewish Health, 2Department of Pharmacology, School of Medicine, University of Colorado Denver The reliability of results in metabolomics experiments depends on the effectiveness and reproducibility of the sample preparation. Described is a rigorous and in-depth method that enables extraction of metabolites from biological fluids with the option of subsequently analyzing up to thousands of compounds, or just the compound classes of interest. Immunology and Infection Non-surgical Intratracheal Instillation of Mice with Analysis of Lungs and Lung Draining Lymph Nodes by Flow Cytometry Manira Rayamajhi1, Elizabeth F. Redente2, Tracy V. Condon1, Mercedes Gonzalez-Juarrero3, David W.H. Riches1,2,4, Laurel L. Lenz1,4 1Department of Immunology, University of Colorado School of Medicine, 2Division of Cell Biology, Department of Pediatrics, National Jewish Health, 3Department of Microbiology, Immunology, and Pathology, Colorado State University, 4Department of Immunology, National Jewish Health We illustrate non-surgical delivery of test materials into the lungs of anesthetized mice via the trachea. This method permits lung exposure to bacterial and viral pathogens, cytokines, antibodies, beads, chemicals, or dyes. We further describe harvesting and processing of lungs and lung draining lymph nodes (LDLNs) for flow cytometry.