Majors Paul D, McLean Jeffrey S, Pinchuk Grigoriy E, Fredrickson James K, Gorby Yuri A, Minard Kevin R, Wind Robert A
Pacific Northwest National Laboratory, MSIN: K8-98, Richland, WA 99352, USA.
J Microbiol Methods. 2005 Sep;62(3):337-44. doi: 10.1016/j.mimet.2005.04.017.
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.
本文描述了用于对活的原位微生物膜进行核磁共振（NMR）光谱和成像研究的新方法及仪器。一个装有平面生物膜支撑体的灌注式NMR/光学显微镜样品室被集成到一个再循环/稀释流动回路生长反应器系统中，并用于原位培养希瓦氏菌属MR-1菌株生物膜。开发了局部NMR技术并用于非侵入性监测时间分辨代谢物浓度以及对生物量体积和分布进行成像。作为该方法可行性的首个例证，进行了一项初始的13C标记乳酸代谢途径研究，所得结果与MR-1的现有基因组数据一致。这些结果代表了朝着我们的最终目标取得的进展，即使用组合的NMR/光学显微镜技术将活的原位生物膜中时间分辨和深度分辨的代谢及质量传输与基因表达相关联。
