title: NMR methods for in situ biofilm metabolism studies.
abstract: Novel procedures and instrumentation are described for nuclear magnetic resonance (NMR) spectroscopy and imaging studies of live, in situ microbial films. A perfused NMR/optical microscope sample chamber containing a planar biofilm support was integrated into a recirculation/dilution flow loop growth reactor system and used to grow in situ Shewanella oneidensis strain MR-1 biofilms. Localized NMR techniques were developed and used to non-invasively monitor time-resolved metabolite concentrations and to image the biomass volume and distribution. As a first illustration of the feasibility of the methodology an initial 13C-labeled lactate metabolic pathway study was performed, yielding results consistent with existing genomic data for MR-1. These results represent progress toward our ultimate goal of correlating time- and depth-resolved metabolism and mass transport with gene expression in live in situ biofilms using combined NMR/optical microscopy techniques.
authors.affiliation: Pacific Northwest National Laboratory, MSIN: K8-98, Richland, WA 99352, USA.
authors.forename: Paul D
authors.initials: PD
authors.lastname: Majors
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: McLean
authors.forename: Grigoriy E
authors.initials: GE
authors.lastname: Pinchuk
authors.forename: James K
authors.initials: JK
authors.lastname: Fredrickson
authors.forename: Yuri A
authors.initials: YA
authors.lastname: Gorby
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Minard
authors.forename: Robert A
authors.initials: RA
authors.lastname: Wind
mesh_terms: D018441:Biofilms / Q000254:growth & development; D019149:Bioreactors; D002247:Carbon Isotopes; D003470:Culture Media; D005650:Fumarates / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000295:instrumentation / Q000379:methods*; D008828:Microbiological Techniques / Q000295:instrumentation; D020592:Shewanella / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; D003470:Culture Media; D005650:Fumarates; D019344:Lactic Acid; C032005:fumaric acid