In this project, we developed a real-time electrochemical mediator assay to enable the assessment of cell numbers and cell viability. It allows us to monitor metabolism calculable down to a single cell in a low cost easy to use rapid assay, as yet not possible with current technology. The developed assay is based on the determination of oxygen. This was made possible via the use of electrochemical mediator ferrocene carboxylic acid (FcA). The FcA showed distinctive catalytic properties in interacting with reactive oxygen species generated from oxygen when compared to ferrocene methanol (FcMeOH). A deeper insight into the chemistry controlling this behaviour is provided. The behaviour is then taken advantage of to develop a cellular aerobic respiration assay. We describe the properties of the FcA system to detect, in real-time, the oxygen consumption of Escherichia coli DH5α (E. coli). We demonstrated that the FcA-based oxygen assay is highly sensitive, and using a population of cells, oxygen consumption rates could be calculated down to a single cell level. More importantly, the results can be accomplished in minutes, considerably outperforming current commercially available biooxygen demand assays. The developed assay is expected to have a significant impact in diverse fields and industries, ranging from environmental toxicology through to pharmaceutical and agrochemical industries. A significant start has been made into optimisation of the system into a commercially marketable product, including much work on mediator immobilisation and electrode synthesis.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:694732 |
| Date | January 2016 |
| Creators | Yip, Nga Chi |
| Publisher | University of Birmingham |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://etheses.bham.ac.uk//id/eprint/6938/ |
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