RESUMO
Nuclear magnetic resonance (NMR) techniques are ideally suited for the study of biofilms and for probing their microenvironments because these techniques allow for noninvasive interrogation and in situ monitoring with high resolution. By combining NMR with simultaneous electrochemical techniques, it is possible to sustain and study live biofilms respiring on electrodes. Here, we describe a biofilm microreactor system, including a reusable and a disposable reactor, that allows for simultaneous electrochemical and NMR techniques (EC-NMR) at the microscale. Microreactors were designed with custom radio frequency resonator coils, which allowed for NMR measurements of biofilms growing on polarized gold electrodes. For an example application of this system we grew Geobacter sulfurreducens biofilms on electrodes. EC-NMR was used to investigate growth medium flow velocities and depth-resolved acetate concentration inside the biofilm. As a novel contribution we used Monte Carlo error analysis to estimate the standard deviations of the acetate concentration measurements. Overall, we found that the disposable EC-NMR microreactor provided a 9.7 times better signal-to-noise ratio over the reusable reactor. The EC-NMR biofilm microreactor system can ultimately be used to correlate extracellular electron transfer rates with metabolic reactions and explore extracellular electron transfer mechanisms.
Assuntos
Biofilmes , Reatores Biológicos , Técnicas Eletroquímicas , Geobacter/fisiologia , Espectroscopia de Ressonância Magnética , MicrofluídicaRESUMO
Nuclear magnetic resonance imaging (NMRI) techniques were employed to identify and selectively image biological films (biofilm) growing in aqueous systems. Biofilms are shown to affect both the longitudinal (T1) and transverse (T2) NMR relaxation time values of proximal water hydrogens. Results are shown for biofilm growth experiments performed in a transparent parallel-plate reactor. A comparison of biofilm distributions by both NMR and optical imaging yielded general agreement for both an open-flow system and an idealized porous system (the reactor without and with packed glass beads, respectively). The selective imaging of biofilm by relaxation NMRI is dependent upon the resolution of relaxation times for the fluid phases, dynamic range, and signal-to-noise ratio. For open-flow systems, the use of a rapid and quantitative T2-sorted NMRI technique was preferred. For porous systems where T2 values are generally more similar, a T1-weighted technique was preferred.
Assuntos
Biofilmes , Espectroscopia de Ressonância Magnética , Reatores Biológicos , Escherichia coli/crescimento & desenvolvimento , Microesferas , PorosidadeRESUMO
Complementary data acquired with different microscopy techniques provide a basis for establishing a more comprehensive understanding of health and disease at a cellular level, particularly when data acquired with different methodologies can be correlated in both time and space. In this Communication, a brief description of a novel instrument capable of simultaneously performing confocal optical and magnetic resonance microscopy is presented, and the first combined images of live Xenopus laevis oocytes are shown. Also, the potential benefits of combined microscopy are discussed, and it is shown that the a priori knowledge of the high-resolution optical images can be used to enhance the boundary resolution and contrast of the MR images.