3 articles published in JoVE
Fluorescence Live-cell Imaging of the Complete Vegetative Cell Cycle of the Slow-growing Social Bacterium Myxococcus xanthus Dominik Schumacher1, Lotte Søgaard-Andersen1 1Department of Ecophysiology, Max Planck Institute for Terrestrial Microbiology Bacterial cells are spatially highly organized. To follow this organization over time in slow growing Myxococcus xanthus cells, a set-up for fluorescence live-cell imaging with high spatiotemporal resolution over several generations was developed. Using this method, spatiotemporal dynamics of important proteins for chromosome segregation and cell division could be determined.
Induction of Cellular Differentiation and Single Cell Imaging of Vibrio parahaemolyticus Swimmer and Swarmer Cells Jan Heering1, Alejandra Alvarado1, Simon Ringgaard1 1Department of Ecophysiology, Max Planck Institute for Terrestrial Microbiology This protocol enables single cell microscopy of the differentially distinct Vibrio parahaemolyticus swimmer and swarmer cells. The method produces a population of swarmer cells easily available for single cell analysis and covers preparation of cell cultures, induction of swarmer differentiation, sample preparation, and image analysis.
Substrate Generation for Endonucleases of CRISPR/Cas Systems Judith Zoephel1, Srivatsa Dwarakanath1, Hagen Richter1, André Plagens1, Lennart Randau1 1Prokaryotic Small RNA Biology, Max-Planck-Institute for Terrestrial Microbiology CRISPR/Cas systems mediate adaptive immunity in Bacteria and Archaea. Many Cas proteins are proposed to act as endoribonucleases acting on crRNA precursors of varying length. Here we illustrate three different approaches to generate pre-crRNA substrates for the biochemical analysis of Cas endonuclease activity.