Methods for Rapid Transfer and Localization of Lyme Disease Pathogens Within the Tick Gut

JoVE 2544 2/14/2011

Toru Kariu¹, Adam S. Coleman¹, John F. Anderson², Utpal Pal¹

¹Department of Veterinary Medicine, University of Maryland, ²Department of Entomology, Connecticut Agricultural Experiment Station

Lyme disease research studies often require generation of ticks infected with the pathogen Borrelia burgdorferi, a process that typically takes several weeks. Here we demonstrate a microinjection-based tick infection procedure that can be accomplished within hours. We also demonstrate an immunofluorescence method for in situ localization of B. burgdorferi within ticks.

Immuno-fluorescence Assay of Leptospiral Surface-exposed Proteins

JoVE 2805 7/01/2011

Marija Pinne^1,2, David Haake^3,4

¹Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, ²Research service, 151, Veterans Affairs Greater Los Angeles Healthcare System, ³Departments of Medicine, Urology at David Geffen School of Medicine and Department of Microbiology, Immunology and Molecular Gentics, University of California Los Angeles (UCLA), ⁴Division of Infectious Diseases, 111F, Veterans Affairs Greater Los Angeles Health Care System

An efficient method to assess surface-exposure of leptospiral proteins is described. The method is specifically designed to avoid disruption of the fragile outer membrane of leptospiral cells. This technique requires employment of several negative controls to assess the integrity of the outer membrane and specificity of antibody reaction.

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

JoVE 3894 2/21/2012

Toni G. Patton¹, Gabrielle Dietrich², Kevin Brandt¹, Marc C. Dolan², Joseph Piesman², Robert D. Gilmore Jr.¹

¹Microbiology and Pathogenesis Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, ²Tick-Borne Diseases Activity, Bacterial Diseases Branch, Division of Vector-Borne Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention

The collection of infected tick hemolymph, salivary glands, and saliva is important to study how tick-borne pathogens cause disease. In this protocol we demonstrate how to collect hemolymph and salivary glands from feeding Ixodes scapularis nymphs. We also demonstrate saliva collection from female I. scapularis adults.

Culturing and Applications of Rotating Wall Vessel Bioreactor Derived 3D Epithelial Cell Models

JoVE 3868 4/03/2012

Andrea L. Radtke, Melissa M. Herbst-Kralovetz

Basic Medical Sciences, University of Arizona College of Medicine - Phoenix

A rotating cell culture system that allows epithelial cells to grow under physiological conditions resulting in 3-D cellular aggregate formation is described. The aggregates generated display in vivo-like characteristics not observed in conventional culture models and serve as a more accurate organotypic model system for a multitude of scientific investigations.

February 2012: This Month in JoVE

JoVE 4258 2/01/2012

Aaron Kolski-Andreaco