PhageFISH for Monitoring Phage Infections at Single Cell Level
PhageFISH uses the power of fluorescence in situ hybridization to monitor intracellular phage infections at single cell level. It combines host cell identification via rRNA probes and phage identification via phage-specific gene probes, allowing for the quantification of the infected cell fraction and the discrimination between infection stages. This book chapter covers all aspects of the procedure, from phage probe design and synthesis, to the phageFISH protocol itself, to microscopy and image analysis.
Key wordsPhageFISH Virus Phage Microorganisms Fluorescence in situ hybridization FISH Infection cycle Infection stages
The authors would like to thank Rudi Amann and Bernhard Fuchs (Max Planck Institute for Marine Microbiology, Bremen) for critical reading of the manuscript and helpful discussions, to Elke Allers (Vibalogics GmbH, Germany), Melissa Duhaime (University of Michigan, USA) and Matthew B. Sullivan (The Ohio State University, USA) for the close partnership which led to the development of the phageFISH method, and finally, to Max Planck Society, Germany, for funding.
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