Studies of the Ecophysiology of Single Cells in Microbial Communities by (Quantitative) Microautoradiography and Fluorescence In Situ Hybridization (MAR-FISH)

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Abstract

Microautoradiography (MAR) in combination with fluorescence in situ hybridization (FISH) is a powerful method of obtaining information about the ecophysiology of probe-defined single cells in mixed microbial communities. The incorporation of radiolabelled substrates can be quantified by automated image analysis (MARQuant). Quantification of MAR signals can answer more specific questions regarding metabolic activity and function of the microbes. Here, we give an overview of how to use MAR-FISH in various ecosystems and provide a detailed protocol for MAR-FISH, including sampling, incubation with radiotracers, the MAR procedure in combination with FISH and other staining techniques, microscopy, and troubleshooting. A description of the MARQuant image analysis tool, including examples of its application, is also provided.
Original languageEnglish
Title of host publicationHydrocarbon and Lipid Microbiology Protocols : Ultrastructure and Imaging
EditorsTerry J. McGenity, Kenneth N. Timmis, Balbina Nogales
PublisherSpringer
Publication date22 Mar 2015
Pages115-130
ISBN (Print)978-3-662-49132-4
ISBN (Electronic)978-3-662-49134-8
DOIs
Publication statusPublished - 22 Mar 2015
SeriesSpringer Protocols Handbooks
ISSN1949-2448

Keywords

  • Microbial communities
  • Radiotracers
  • Ecophysiology
  • FISH
  • Microautoradiography

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