NMR methods for in-situ biofilm metabolism studies : spatial and temporal resolved measurements

• Published in: Water Science & Technology Vol. 52; no. 7; pp. 7 - 12
• Main Authors: MAJORS, P. D, MCLEAN, J. S, FREDRICRSON, J. K, WIND, R. A
• Format: Conference Proceeding Journal Article
• Language: English
• Published: Oxford Pergamon 01.10.2005; IWA Publishing
• Subjects: Adherent cells; Analytical methods; Applied sciences; Biofilms; Biological and medical sciences; Biological treatment of waters; Biotechnology; Environment and pollution; Exact sciences and technology; Fluorescence; Fundamental and applied biological sciences. Psychology; Gene expression; General purification processes; Growth conditions; Industrial applications and implications. Economical aspects; Light microscopy; Magnetic resonance imaging; Mass transport; Metabolic flux; Metabolic pathways; Metabolism; Metabolites; Microscopy; NMR; Nuclear magnetic resonance; Optical microscopy; Optics; Pollution; Shewanella oneidensis; Spatial discrimination; Spatial resolution; Spectroscopy; Wastewaters; Water treatment and pollution
• ISBN: 9781843395522; 1843395525
• ISSN: 0273-1223; 1996-9732
• DOI: 10.2166/wst.2005.0174

We are developing novel nuclear magnetic resonance (NMR) microscopy, spectroscopy and combined NMR/optical techniques for the study of biofilms under known, controlled growth conditions. Objectives include: time and depth-resolved metabolite concentrations with isotropic spatial resolution on the order of 10 microns, metabolic pathways and flux rates, mass transport and ultimately their correlation with gene expression by optical microscopy in biofilms. We describe the implementation of ex-situ grown biofilms to improve growth environment control and NMR analysis. In-situ NMR depth resolved metabolite profiling techniques are introduced and demonstrated for a Shewanella oneidensis strain MR-1 biofilm. Finally, initial combined confocal fluorescence and magnetic resonance images are shown for a GFP-labeled Shewanella biofilm. These methods are equally applicable to other biofilm systems of interest; thus they may provide a significant contribution toward the understanding of adherent cell metabolism.