• Open Access

Simultaneous analysis of microbial identity and function using NanoSIMS

Authors

  • Tianlun Li,

    1. Cemagref, Unité de recherche Hydrosystèmes et bioprocédés, Parc de Tourvoie, BP 44, 92163 Antony cedex, France.
    2. Université Paris-Sud, Institut de Génétique et Microbiologie, Orsay, 91405 France.
    Search for more papers by this author
  • Ting-Di Wu,

    1. Institut Curie, Laboratoire de Microscopie Ionique, Orsay, 91405 France.
    2. INSERM, U759, Orsay, 91405 France.
    Search for more papers by this author
  • Laurent Mazéas,

    Corresponding author
    1. Cemagref, Unité de recherche Hydrosystèmes et bioprocédés, Parc de Tourvoie, BP 44, 92163 Antony cedex, France.
    Search for more papers by this author
  • Laurent Toffin,

    1. Cemagref, Unité de recherche Hydrosystèmes et bioprocédés, Parc de Tourvoie, BP 44, 92163 Antony cedex, France.
    2. Laboratoire de Microbiologie des Environnements Extrêmes (UMR6197) IFREMER Centre de Brest, Technopôle Brest-Iroise, BP70, 29280 Plouzané, France.
    Search for more papers by this author
  • Jean-Luc Guerquin-Kern,

    1. Institut Curie, Laboratoire de Microscopie Ionique, Orsay, 91405 France.
    2. INSERM, U759, Orsay, 91405 France.
    Search for more papers by this author
  • Gérard Leblon,

    1. Université Paris-Sud, Institut de Génétique et Microbiologie, Orsay, 91405 France.
    Search for more papers by this author
  • Théodore Bouchez

    Corresponding author
    1. Cemagref, Unité de recherche Hydrosystèmes et bioprocédés, Parc de Tourvoie, BP 44, 92163 Antony cedex, France.
    Search for more papers by this author

  • Re-use of this article is permitted in accordance with the Creative, Commons Deed, Attribution 2.5, which does not permit commercial exploitation.

*E-mail theodore.bouchez@cemagref.fr; laurent.mazeas@cemagref.fr; Tel. (+33) 140966040; Fax (+33) 140966072.

Summary

Identifying the function of uncultured microbes in their environments today remains one of the main challenges for microbial ecologists. In this article, we describe a new method allowing simultaneous analysis of microbial identity and function. This method is based on the visualization of oligonucleotide probe-conferred hybridization signal in single microbial cells and isotopic measurement using high-resolution ion microprobe (NanoSIMS). In order to characterize the potential of the method, an oligonucleotide containing iodized cytidine was hybridized on fixed cells of Escherichia coli cultured on media containing different levels of 13C or 15N. Iodine signals could clearly be localized on targeted cells and the isotopic enrichment could be monitored at the single-cell level. The applicability of this new technique to the study of in situ ecophysiology of uncultured microorganisms within complex microbial communities is illustrated.

Ancillary