This material was originally published in the Purdue Cytometry CD-ROM Series

Quantitative flow cytometric detection of specific microorganisms in soil samples.

Jean-Christophe Thomas (1), Marcel Desrosiers (2), Richard Villemur (3) and Yves St-Pierre (2)

Specific detection and accurate enumeration of microbial populations in the environment is crucial during bioremediation treatments, in which microorganisms are inoculated in soil to breakdown pollutants. Up to now, enumeration of specific microorganisms in environmental samples relied on counting visible colonies on selective agar media (CFU) or on epifluorescence microscopic counting.

Flow cytometry (FCM) offers the ability to perform rapid, multiparametric analysis of specific individual cells in a heterogeneous population. However, the use of FCM to monitor bacteria in soil has been hampered by the high levels of autofluorescence from environmental samples and by background noise due to non-specific attachment of the fluorescent probes to debris.

In this work, we report a new multiparametric strategy that allows specific and quantitative detection of Sphingomonas sp. strain 107 inoculated into soil samples. This method was developed in five successive steps:

(1) Ecologie Microbienne du Sol
UMR CNRS 5557, Universite Lyon 1
La Doua, Bat. 741
43 bd du 11 Novembre 1918
69622 Villeurbanne Cedex, France

(2) Centre de Recherche en Immunologie
Institut Armand-Frappier
531 Boul. des Prairies
Laval (Quebec), Canada H7V 1B7
email: yves_st-pierre@iaf.UQUEBEC.CA

(3) Centre de Recherche en Microbiologie Appliquee
Institut Armand-Frappier
531 Boul. des Prairies
Laval (Quebec), Canada H7V 1B7.CA
email: richard_villemur@iaf.UQUEBEC.CA

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