Journal article
Bioelectrochemical probing of intracellular redox processes in living yeast cells—application of redox polymer wiring in a microfluidic environment
Department of Micro- and Nanotechnology, Technical University of Denmark1
Bioanalytics, Department of Micro- and Nanotechnology, Technical University of Denmark2
Nanointegration, Department of Micro- and Nanotechnology, Technical University of Denmark3
University of Canterbury4
Lund University5
Fluidic Array Systems and Technology, Department of Micro- and Nanotechnology, Technical University of Denmark6
Conventionally, microbial bioelectrochemical assays have been conducted using immobilized cells on an electrode that is placed in an electrochemical batch cell. In this paper, we describe a developed microfluidic platform with integrated microelectrode arrays for automated bioelectrochemical assays utilizing a new double mediator system to map redox metabolism and screen for genetic modifications in Saccharomyces cerevisiae cells.
The function of this new double mediator system based on menadione and osmium redox polymer (PVI-Os) is demonstrated. “Wiring” of S. cerevisiae cells using PVI-Os shows a significant improvement of bioelectrochemical monitoring in a microfluidic environment and functions as an effective immobilization matrix for cells that are not strongly adherent.
The function of the developed microfluidic platform is demonstrated using two strains of S. cerevisiae, ENY.WA and its deletion mutant EBY44, which lacks the enzyme phosphoglucose isomerase. The cellular responses to introduced glucose and fructose were recorded for the two S. cerevisiae strains, and the obtained results are compared with previously published work when using an electrochemical batch cell, indicating that microfluidic bioelectrochemical assays employing the menadione–PVI-Os double mediator system provides an effective means to conduct automated microbial assays.
FigureMicrofluidic platform for bioelectrochemical assays using osmium redox polymer “wired” living yeast cells
| Language: | English |
|---|---|
| Publisher: | Springer-Verlag |
| Year: | 2013 |
| Pages: | 3847-3858 |
| ISSN: | 16182650 and 16182642 |
| Types: | Journal article |
| DOI: | 10.1007/s00216-013-6709-4 |
| ORCIDs: | Heiskanen, Arto , Dufva, Martin and Emnéus, Jenny |
Cellular redox activity Microbial bioelectrochemistry Microfluidic system Osmium redox polymer Osmium redox polymer system Saccharomyces cerevisiae
Analytical Chemistry Biochemistry, general Cells, Immobilized Characterization and Evaluation of Materials Chemistry Chemistry and Materials Science Double mediator system Electrochemical Techniques Environmental Monitoring/Analysis Equipment Design Food Science Fructose Gene Deletion Glucose Laboratory Medicine Microelectrodes Microfluidics Osmium Oxidation-Reduction Polymers Vitamin K 3
