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John Innes Centre, Norwich Research Park

2 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Fluorescence Time-lapse Imaging of the Complete S. venezuelae Life Cycle Using a Microfluidic Device

Susan Schlimpert¹, Klas Flärdh², Mark J. Buttner¹

¹Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, ²Department of Biology, Lund University

Streptomyces are characterized by a complex life cycle that has been experimentally challenging to study by cell biological means. Here we present a protocol to perform fluorescence time-lapse microscopy of the complete life cycle by growing Streptomyces venezuelae in a microfluidic device.

Biology

Real-time In Vivo Recording of Arabidopsis Calcium Signals During Insect Feeding Using a Fluorescent Biosensor

Thomas R. Vincent¹, James Canham¹, Masatsugu Toyota^2,3,4, Marieta Avramova¹, Sam T. Mugford⁵, Simon Gilroy², Anthony J. Miller¹, Saskia Hogenhout⁵, Dale Sanders¹

¹Department of Metabolic Biology, John Innes Centre, Norwich Research Park, ²Department of Botany, University of Wisconsin, Madison, ³Department of Biochemistry and Molecular Biology, Saitama University, ⁴Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), ⁵Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park

This protocol outlines a simple method for analyzing calcium signals in plants generated by feeding hemipteran insects, such as aphids. Arabidopsis thaliana transformed with the GFP calcium biosensor GCaMP3 allow for the real-time in vivo imaging of calcium dynamics with a high temporal and spatial resolution.