3 articles published in JoVE
Human Lung Dendritic Cells: Spatial Distribution and Phenotypic Identification in Endobronchial Biopsies Using Immunohistochemistry and Flow Cytometry Faezzah Baharom1, Gregory Rankin2, Saskia Scholz1, Jamshid Pourazar2, Clas Ahlm3, Anders Blomberg2, Anna Smed-Sörensen1 1Immunology and Allergy Unit, Department of Medicine Solna, Karolinska Institutet, 2Division of Medicine, Department of Public Health and Clinical Medicine, Umeå University, 3Divison of Infectious Diseases, Department of Clinical Microbiology, Umeå University Lung-resident immune cells, including dendritic cells (DCs) in humans, are critical for defense against inhaled pathogens and allergens. However, due to the scarcity of human lung tissue, studies are limited. This work presents protocols to process human mucosal endobronchial biopsies for studying lung DCs using immunohistochemistry and flow cytometry.
Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography Samantha M. Desmarais1, Felipe Cava2, Miguel A. de Pedro3, Kerwyn Casey Huang1,4 1Department of Bioengineering, Stanford University, 2Department of Molecular Biology and Laboratory for Molecular Infection Medicine Sweden, Umeå Centre for Microbial Research, Umeå University, 3Campus de Cantoblanco, Universidad Autonoma de Madrid, 4Department of Microbiology and Immunology, Stanford University School of Medicine The bacterial cell wall is composed of peptidoglycan, a macromolecular network of sugar strands crosslinked by peptides. Ultra Performance Liquid Chromatography provides high resolution and throughput for novel discoveries of peptidoglycan composition. We present a procedure for the isolation of cell walls (sacculi) and their subsequent preparation for analysis via UPLC.
Near Infrared Optical Projection Tomography for Assessments of β-cell Mass Distribution in Diabetes Research Anna U. Eriksson*1, Christoffer Svensson*1, Andreas Hörnblad1, Abbas Cheddad1, Elena Kostromina1, Maria Eriksson1, Nils Norlin1, Antonello Pileggi2, James Sharpe3, Fredrik Georgsson4, Tomas Alanentalo1, Ulf Ahlgren1 1Umeå Centre for Molecular Medicine, Umeå University, 2Cell Transplant Center, Diabetes Research Institute, University of Miami,, 3EMBL-CRG Systems Biology Program, Centre for Genomic Regulation, Catalan Institute of Research and Advanced Studies, 4Dept. of Computing Science, Umeå University We describe the adaptation of optical projection tomography (OPT)1 to imaging in the near infrared spectrum, and the implementation of a number of computational tools. These protocols enable assessments of pancreatic β-cell mass (BCM) in larger specimens, increase the multichannel capacity of the technique and increase the quality of OPT data.