University of East Anglia 5 articles published in JoVE Immunology and Infection An Intravital Microscopy-Based Approach to Assess Intestinal Permeability and Epithelial Cell Shedding Performance Luz DC. Martínez-Sánchez*1, Rashmita Pradhan*1, Phuong A. Ngo1, Lena Erkert1, Lukas S. Becker1, Alastair J. Watson2, Imke Atreya*1, Markus F. Neurath*1, Rocío López-Posadas*1 1Medical Clinic 1, University of Erlangen-Nuremberg, University Hospital of Erlangen, 2Norwich Medical School, University of East Anglia, Norwich Research Park Taking advantage of intravital microscopy, the method presented here enables real-time visualization of intestinal epithelial cell shedding in living animals. Therefore, topically stained intestinal mucosa (acriflavine and rhodamineB-dextran) of anesthetized mice is imaged up to single-cell resolution using confocal microscopy. Behavior The (Spatial) Memory Game: Testing the Relationship Between Spatial Language, Object Knowledge, and Spatial Cognition Harmen B. Gudde1, Debra Griffiths1, Kenny R. Coventry1 1School of Psychology, University of East Anglia We present a protocol to explore the relationship between spatial language production, spatial memory, and object knowledge. The procedure allows experimental manipulation of, and control over, conditions of object knowledge, language at instruction, and physical location, thus teasing apart cognitive and linguistic models describing interactions between these variables. Developmental Biology Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids Deborah A. Goldspink1, Zoe J. Matthews2, Elizabeth K. Lund1, Tom Wileman2, Mette M. Mogensen1 1School of Biological Sciences, University of East Anglia, 2Norwich Medical School, University of East Anglia We present protocols for isolation of intestinal 3D structures from in vivo tissue and in vitro basement matrix embedded organoids, and detail different fixation and staining protocols optimized for immuno-labeling of microtubule, centrosomal, and junctional proteins as well as cell markers including the stem cell protein Lgr5. Chemistry Spin Saturation Transfer Difference NMR (SSTD NMR): A New Tool to Obtain Kinetic Parameters of Chemical Exchange Processes María Teresa Quirós1, Colin Macdonald1,2, Jesús Angulo2, María Paz Muñoz1 1School of Chemistry, University of East Anglia, 2School of Pharmacy, University of East Anglia A detailed protocol describing the SSTD NMR method is presented here to help new users apply this new method to obtain the kinetic parameters of their own systems undergoing chemical exchange. Biochemistry Species Determination and Quantitation in Mixtures Using MRM Mass Spectrometry of Peptides Applied to Meat Authentication Yvonne Gunning1, Andrew D. Watson1, Neil M. Rigby2, Mark Philo1, Joshua K. Peazer1,3, E. Kate Kemsley1 1Analytical Sciences Unit, Institute of Food Research, 2Institute of Food Research, 3School of Chemistry, University of East Anglia We present a protocol for identifying and quantifying the components in mixtures of species possessing similar proteins. Mass spectrometry detects peptides for identification, and gives relative quantitation by ratios of peak areas. As a tool food for fraud detection, the method can detect 1% horse in beef.
