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A plate-based electrochromic approach for the high-throughput detection of electrochemically active bacteria

Nature Protocols volume 9, pages 112–119 (2014)Cite this article

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Abstract

Electrochemically active bacteria (EAB) have the ability to transfer electrons to electron acceptors located outside the cell, and they are widely present in diverse environments. In spite of their important roles in geochemical cycles, environmental remediation and electricity generation, so far, only a limited number and types of EAB have been isolated and characterized. Thus, effective and rapid EAB identification methods are highly desirable. In this protocol, we describe a photometric protocol for the visualization and high-throughput identification and isolation of EAB. The protocol relies on the fast electron acquisition and color change ability of an electrochromic material, namely a tungsten trioxide (WO3) nanorod assembly. The extracellular electron transfer (EET) from EAB to the WO3 nanorod assembly probe is accompanied by a bioelectrochromic reaction made evident by the color change of the probe. This protocol enables researchers to rapidly identify EAB and evaluate their EET ability either qualitatively with the naked eye or quantitatively by image analysis. We have also successfully used this protocol to isolate EAB from environmental samples. The time needed to complete this protocol is ∼2 d, with the actual EAB identification process taking about 5 min.

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Figure 1: Workflow of EAB identification.
Figure 2: An illustration of the EAB colony isolation process.
Figure 3: Correlation between the population density of cells and the chromaticity.
Figure 4: Screenshot of the 'Density (mean)' measurement process.

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Acknowledgements

The authors thank the National Nature Science Foundation of China (51129803) and the Program for Changjiang Scholars and Innovative Research Team in University, China for the partial support of this study. They also thank Professor K.H. Nealson from the University of Southern California for kindly providing S. oneidensis MR-1 wild type and its mutant strains.

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  1. Shi-Jie Yuan and Wen-Wei Li: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, University of Science and Technology of China, Hefei, China

    Shi-Jie Yuan, Wen-Wei Li, Yuan-Yuan Cheng, Hui He, Jie-Jie Chen, Zhong-Hua Tong, Zhi-Qi Lin, Feng Zhang, Guo-Ping Sheng & Han-Qing Yu

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Contributions

S.-J.Y., W.-W.L., G.-P.S., Z.-H.T. and H.-Q.Y. designed the experiments; S.-J.Y., W.-W.L., Y.-Y.C., H.H., J.-J.C., Z.-Q.L. and F.Z. conducted the experiments; S.-J.Y., W.-W.L., G.-P.S., Y.-Y.C., F.Z., Z.-H.T., J.-J.C. and H.-Q.Y. wrote and edited the manuscript. All authors contributed to discussion about the results and implications of the manuscript.

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Correspondence to Guo-Ping Sheng or Han-Qing Yu.

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The authors declare no competing financial interests.

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Yuan, SJ., Li, WW., Cheng, YY. et al. A plate-based electrochromic approach for the high-throughput detection of electrochemically active bacteria. Nat Protoc 9, 112–119 (2014). https://doi.org/10.1038/nprot.2013.173

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