PhageFISH for Monitoring Phage Infections at Single Cell Level

Jimena Barrero-Canosa; Cristina Moraru

doi:10.1007/978-1-4939-8940-9_1

PhageFISH for Monitoring Phage Infections at Single Cell Level

Methods Mol Biol. 2019:1898:1-26. doi: 10.1007/978-1-4939-8940-9_1.

Authors

Jimena Barrero-Canosa¹, Cristina Moraru²

Affiliations

¹ Department of Environmental Technology., Technische Universität Berlin, Berlin, Germany.
² Institute for Chemistry and Biology of the Marine Environment, Oldenburg, Germany. liliana.cristina.moraru@uni-oldenburg.de.

PMID: 30570719
DOI: 10.1007/978-1-4939-8940-9_1

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.

Keywords: FISH; Fluorescence in situ hybridization; Infection cycle; Infection stages; Microorganisms; Phage; PhageFISH; Virus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacteriophages / growth & development*
Bacteriophages / pathogenicity
In Situ Hybridization, Fluorescence / methods*
Microscopy / methods
Oligonucleotide Probes / genetics
Single-Cell Analysis / methods*

Substances

Oligonucleotide Probes