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Rapid resonance Raman microspectroscopy to probe carbon dioxide fixation by single cells in microbial communities

Lookup NU author(s): Professor Grant Burgess


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Photosynthetic microorganisms play crucial roles in aquatic ecosystems and are the major primary producers in global marine ecosystems. The discovery of new bacteria and microalgae that play key roles in CO2 fixation is hampered by the lack of methods to identify hitherto-unculturable microorganisms. To overcome this problem we studied single microbial cells using stable-isotope probing (SIP) together with resonance Raman (RR) microspectroscopy of carotenoids, the light-absorbing pigments present in most photosynthetic microorganisms. We show that fixation of (CO2)-C-13 into carotenoids produces a red shift in single-cell RR (SCRR) spectra and that this SCRR-SIP technique is sufficiently sensitive to detect as little as 10% of C-13 incorporation. Mass spectrometry (MS) analysis of labelled cellular proteins verifies that the red shift in carotenoid SCRR spectra acts as a reporter of the C-13 content of single cells. Millisecond Raman imaging of cells in mixed cultures and natural seawater samples was used to identify cells actively fixing CO2, demonstrating that the SCRR-SIP is a noninvasive method for the rapid and quantitative detection of CO2 fixation at the single cell level in a microbial community. The SCRR-SIP technique may provide a direct method for screening environmental samples, and could help to reveal the ecophysiology of hitherto-unculturable microorganisms, linking microbial species to their ecological function in the natural environment. The ISME Journal (2012) 6, 875-885; doi: 10.1038/ismej.2011.150; published online 24 November 2011

Publication metadata

Author(s): Li MQ, Canniffe DP, Jackson PJ, Davison PA, FitzGerald S, Dickman MJ, Burgess JG, Hunter CN, Huang WE

Publication type: Article

Publication status: Published

Journal: ISME Journal

Year: 2012

Volume: 6

Issue: 4

Pages: 875-885

Print publication date: 24/11/2011

ISSN (print): 1751-7362

ISSN (electronic): 1751-7370

Publisher: Nature Publishing Group


DOI: 10.1038/ismej.2011.150


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