title: Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach.
abstract: The wide collection of currently available fluorescent proteins (FPs) offers new possibilities for multicolor reporter gene-based studies of bacterial functions. However, the simultaneous use of multiple FPs is often limited by the bleed-through of their emission spectra. Here we introduce an original approach for detection and separation of multiple overlapping fluorescent signals from mixtures of bioreporters strains. The proposed method relies on the coupling of synchronous fluorescent spectroscopy (SFS) with blind spectral decomposition achieved by the Canonical Polyadic (CP) decomposition (also known as Candecomp/Parafac) of three-dimensional data arrays. Due to the substantial narrowing of FP emission spectra and sensitive detection of multiple FPs in a one-step scan, SFS reduced spectral overlap and improved the selectivity of the CP unmixing procedure. When tested on mixtures of labeled E. coli strains, the SFS/CP approach could easily extract the contribution of at least four overlapping FPs. Furthermore, it allowed to simultaneously monitor the expression of three iron responsive genes and pyoverdine production in P. aeruginosa. Implemented in a convenient microplate format, this multiplex fluorescent reporter method provides a useful tool to study complex processes with different variables in bacterial systems.
authors.affiliation: Université de Lorraine, CNRS, Laboratoire Interdisciplinaire des Environnements Continentaux (LIEC), UMR 7360, Vandoeuvre-lès-Nancy, France.
authors.forename: Damien
authors.initials: D
authors.lastname: Parrello
authors.affiliation: Université de Lorraine, CNRS, Laboratoire Interdisciplinaire des Environnements Continentaux (LIEC), UMR 7360, Vandoeuvre-lès-Nancy, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Mustin
authors.affiliation: Université de Lorraine, CNRS, Centre de Recherche en Automatique de Nancy (CRAN) UMR 7039, Vandoeuvre-lès-Nancy, France.
authors.forename: David
authors.initials: D
authors.lastname: Brie
authors.affiliation: Université de Lorraine, CNRS, Centre de Recherche en Automatique de Nancy (CRAN) UMR 7039, Vandoeuvre-lès-Nancy, France.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Miron
authors.affiliation: Université de Lorraine, CNRS, Laboratoire Interdisciplinaire des Environnements Continentaux (LIEC), UMR 7360, Vandoeuvre-lès-Nancy, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Billard
mesh_terms: D003116:Color; D004926:Escherichia coli / Q000235:genetics*; D006706:Homeostasis; D007501:Iron / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D008164:Luminescent Proteins; D007501:Iron
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