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PhageFISH for Monitoring Phage Infections at Single Cell Level

  • Jimena Barrero-Canosa
  • Cristina MoraruEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1898)

Abstract

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 words

PhageFISH Virus Phage Microorganisms Fluorescence in situ hybridization FISH Infection cycle Infection stages 

Notes

Acknowledgments

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute for Chemistry and Biology of the Marine EnvironmentOldenburgGermany
  2. 2.Department of Environmental Technology.Technische Universität BerlinBerlinGermany

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