Wageningen University

5 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Avian Influenza Surveillance with FTA Cards: Field Methods, Biosafety, and Transportation Issues Solved

Robert H.S. Kraus¹, Pim van Hooft¹, Jonas Waldenström², Neus Latorre-Margalef², Ronald C. Ydenberg^1,3, Herbert H.T. Prins¹

¹Resource Ecology Group, Wageningen University, ²Section for Zoonotic Ecology and Epidemiology, School of Natural Sciences, Linnaeus University, ³Centre for Wildlife Ecology, Simon Fraser University

A method to preserve, detect and sequence RNA from Avian Influenza Viruses was validated and extended using natural faecal samples from birds. This technique removes the necessity of maintaining a cool chain and handling of infectious viruses and can be applied in a 96-well high-throughput setup.

Biology

Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana

Juan Du^1,2, Hendrik Rietman¹, Vivianne G. A. A. Vleeshouwers¹

¹Wageningen UR Plant Breeding, Wageningen University, ²Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University, Ministry of Education, National Centre for Vegetable Improvement (Central China), Potato Engineering and Technology Research Centre of Hubei Province, Huazhong Agricultural University

Agroinfiltration and PVX agroinfection are routine functional assays for transient ectopic expression of genes in plants. These methods are efficient assays in effectoromics strategies (rapid resistance and avirulence gene discovery) and crucial to modern research in molecular plant pathology. They meet the demand for robust high-throughput functional analysis in plants.

Environment

Modification and Application of a Leaf Blower-vac for Field Sampling of Arthropods

Yi Zou¹, Mario D. van Telgen¹, Junhui Chen², Haijun Xiao², Joop de Kraker³, Felix J. J. A. Bianchi⁴, Wopke van der Werf¹

¹Centre for Crop Systems Analysis, Wageningen University, ²Institute of Entomology, Jiangxi Agricultural University, ³Department of Science, Open University, ⁴Farming Systems Ecology, Wageningen University

Assessment of arthropod abundance in crops is critical for investigating population dynamics and species interactions. Here we describe the modification and application of a leaf blower-vac for suction sampling of arthropods in rice.

Cancer Research

Detection of Mitochondria Membrane Potential to Study CLIC4 Knockdown-induced HN4 Cell Apoptosis In Vitro

Jinsen Lu¹, Lele Wu², Xiaoke Wang², Jinhang Zhu², Juan Du², Bing Shen²

¹School of Basic Medical Sciences, Anhui Medical University, ²Department of Orthopedics, Anhui Provincial Hospital, Anhui Medical University

Here we present a detailed protocol for the application of rhodamine 123 to identify the mitochondrial membrane potential (MMP) and study CLIC4 knockdown-induced HN4 cell apoptosis in vitro. Under common fluorescence microscope and confocal laser scanning fluorescence microscope, the real-time change of the MMP was recorded.

Biochemistry

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy

Emery Haley^*1, Wooyoung Choi^*1, Chen Fan¹, Weinan Sun^2,3, Juan Du¹, Wei Lü¹

¹Van Andel Institute, ²Vollum Institute, ³Janelia Research Campus

This protocol describes techniques used to determine ion channel structures by cryo-electron microscopy, including a baculovirus system used to efficiently express genes in mammalian cells with minimum effort and toxicity, protein extraction, purification, and quality checking, sample grid preparation and screening, as well as data collection and processing.