North Carolina State University College of Veterinary Medicine 2 articles published in JoVE Biology Sequential Immunofluorescence and Immunohistochemistry on Cryosectioned Zebrafish Embryos Jordan L. Ferguson, Heather R. Shive 1Department of Population Health and Pathobiology, NC State University College of Veterinary Medicine This protocol demonstrates sequential immunofluorescence and immunohistochemistry on cryosections from early-stage zebrafish embryos enabling precise colocalization analyses in specific cell populations. Immunology and Infection Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291 Jenessa A. Winston1, Rajani Thanissery1, Stephanie A. Montgomery2, Casey M. Theriot1 1Department of Population Health and Pathobiology, North Carolina State University College of Veterinary Medicine, 2Department of Pathology and Laboratory Medicine, Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine This protocol outlines the cefoperazone mouse model of Clostridium difficile infection (CDI) using a clinically-relevant and genetically-tractable strain, R20291. Emphasis on clinical disease monitoring, C. difficile bacterial enumeration, toxin cytotoxicity, and histopathological changes throughout CDI in a mouse model are detailed in the protocol.