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Integrated circuit-based electrochemical sensor for spatially resolved detection of redox-active metabolites in biofilms.

ABSTRACT: Despite advances in monitoring spatiotemporal expression patterns of genes and proteins with fluorescent probes, direct detection of metabolites and small molecules remains challenging. A technique for spatially resolved detection of small molecules would benefit the study of redox-active metabolites that are produced by microbial biofilms and can affect their development. Here we present an integrated circuit-based electrochemical sensing platform featuring an array of working electrodes and parallel potentiostat channels. 'Images' over a 3.25 × 0.9?mm(2) area can be captured with a diffusion-limited spatial resolution of 750??m. We demonstrate that square wave voltammetry can be used to detect, identify and quantify (for concentrations as low as 2.6??M) four distinct redox-active metabolites called phenazines. We characterize phenazine production in both wild-type and mutant Pseudomonas aeruginosa PA14 colony biofilms, and find correlations with fluorescent reporter imaging of phenazine biosynthetic gene expression.

SUBMITTER: Bellin DL

PROVIDER: S-EPMC3969851 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Integrated circuit-based electrochemical sensor for spatially resolved detection of redox-active metabolites in biofilms.

Bellin Daniel L DL Sakhtah Hassan H Rosenstein Jacob K JK Levine Peter M PM Thimot Jordan J Emmett Kevin K Dietrich Lars E P LE Shepard Kenneth L KL

Nature communications 20140101

Despite advances in monitoring spatiotemporal expression patterns of genes and proteins with fluorescent probes, direct detection of metabolites and small molecules remains challenging. A technique for spatially resolved detection of small molecules would benefit the study of redox-active metabolites that are produced by microbial biofilms and can affect their development. Here we present an integrated circuit-based electrochemical sensing platform featuring an array of working electrodes and pa ...[more]

PMID: 24510163

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Project description:BackgroundSimultaneous optical mapping of multiple electrophysiologically relevant parameters in living myocardium is desirable for integrative exploration of mechanisms underlying heart rhythm generation under normal and pathophysiologic conditions. Current multiparametric methods are technically challenging, usually involving multiple sensors and moving parts, which contributes to high logistic and economic thresholds that prevent easy application of the technique.ObjectiveThe purpose of this study was to develop a simple, affordable, and effective method for spatially resolved, continuous, simultaneous, and multiparametric optical mapping of the heart, using a single camera.MethodsWe present a new method to simultaneously monitor multiple parameters using inexpensive off-the-shelf electronic components and no moving parts. The system comprises a single camera, commercially available optical filters, and light-emitting diodes (LEDs), integrated via microcontroller-based electronics for frame-accurate illumination of the tissue. For proof of principle, we illustrate measurement of four parameters, suitable for ratiometric mapping of membrane potential (di-4-ANBDQPQ) and intracellular free calcium (fura-2), in an isolated Langendorff-perfused rat heart during sinus rhythm and ectopy, induced by local electrical or mechanical stimulation.ResultsThe pilot application demonstrates suitability of this imaging approach for heart rhythm research in the isolated heart. In addition, locally induced excitation, whether stimulated electrically or mechanically, gives rise to similar ventricular propagation patterns.ConclusionCombining an affordable camera with suitable optical filters and microprocessor-controlled LEDs, single-sensor multiparametric optical mapping can be practically implemented in a simple yet powerful configuration and applied to heart rhythm research. The moderate system complexity and component cost is destined to lower the threshold to broader application of functional imaging and to ease implementation of more complex optical mapping approaches, such as multiparametric panoramic imaging. A proof-of-principle application confirmed that although electrically and mechanically induced excitation occur by different mechanisms, their electrophysiologic consequences downstream from the point of activation are not dissimilar.

Dataset Information

Integrated circuit-based electrochemical sensor for spatially resolved detection of redox-active metabolites in biofilms.

Publications

Integrated circuit-based electrochemical sensor for spatially resolved detection of redox-active metabolites in biofilms.

Similar Datasets

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