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

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Resumé

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.
OriginalsprogEngelsk
TitelHydrocarbon and Lipid Microbiology Protocols : Ultrastructure and Imaging
RedaktørerTerry J. McGenity, Kenneth N. Timmis, Balbina Nogales
ForlagSpringer
Publikationsdato22 mar. 2015
Sider115-130
ISBN (Trykt)978-3-662-49132-4
ISBN (Elektronisk)978-3-662-49134-8
DOI
StatusUdgivet - 22 mar. 2015
NavnSpringer Protocols Handbooks
ISSN1949-2448

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Fluorescence
Image analysis
Ecosystems
Microscopic examination
Sampling
Substrates

Citer dette

Nierychlo, M., Nielsen, J. L., & Nielsen, P. H. (2015). Studies of the Ecophysiology of Single Cells in Microbial Communities by (Quantitative) Microautoradiography and Fluorescence In Situ Hybridization (MAR-FISH). I T. J. McGenity, K. N. Timmis, & B. Nogales (red.), Hydrocarbon and Lipid Microbiology Protocols: Ultrastructure and Imaging (s. 115-130). Springer. Springer Protocols Handbooks https://doi.org/10.1007/8623_2015_66
Nierychlo, Marta ; Nielsen, Jeppe Lund ; Nielsen, Per Halkjær. / Studies of the Ecophysiology of Single Cells in Microbial Communities by (Quantitative) Microautoradiography and Fluorescence In Situ Hybridization (MAR-FISH). Hydrocarbon and Lipid Microbiology Protocols: Ultrastructure and Imaging. red. / Terry J. McGenity ; Kenneth N. Timmis ; Balbina Nogales. Springer, 2015. s. 115-130 (Springer Protocols Handbooks).
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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.",
keywords = "Microbial communities, Radiotracers, Ecophysiology, FISH, Microautoradiography",
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Nierychlo, M, Nielsen, JL & Nielsen, PH 2015, Studies of the Ecophysiology of Single Cells in Microbial Communities by (Quantitative) Microautoradiography and Fluorescence In Situ Hybridization (MAR-FISH). i TJ McGenity, KN Timmis & B Nogales (red), Hydrocarbon and Lipid Microbiology Protocols: Ultrastructure and Imaging. Springer, Springer Protocols Handbooks, s. 115-130. https://doi.org/10.1007/8623_2015_66

Studies of the Ecophysiology of Single Cells in Microbial Communities by (Quantitative) Microautoradiography and Fluorescence In Situ Hybridization (MAR-FISH). / Nierychlo, Marta; Nielsen, Jeppe Lund; Nielsen, Per Halkjær.

Hydrocarbon and Lipid Microbiology Protocols: Ultrastructure and Imaging. red. / Terry J. McGenity; Kenneth N. Timmis; Balbina Nogales. Springer, 2015. s. 115-130 (Springer Protocols Handbooks).

Publikation: Bidrag til bog/antologi/rapport/konference proceedingBidrag til bog/antologiForskningpeer review

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T1 - Studies of the Ecophysiology of Single Cells in Microbial Communities by (Quantitative) Microautoradiography and Fluorescence In Situ Hybridization (MAR-FISH)

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N2 - 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.

AB - 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.

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Nierychlo M, Nielsen JL, Nielsen PH. Studies of the Ecophysiology of Single Cells in Microbial Communities by (Quantitative) Microautoradiography and Fluorescence In Situ Hybridization (MAR-FISH). I McGenity TJ, Timmis KN, Nogales B, red., Hydrocarbon and Lipid Microbiology Protocols: Ultrastructure and Imaging. Springer. 2015. s. 115-130. (Springer Protocols Handbooks). https://doi.org/10.1007/8623_2015_66