Development of a four electrode sensor array for impedance spectroscopy in high content screenings of fermentation processes

Abstract There is an increasing demand of efficient screening techniques with high information output in biotechnological research and pharmaceutical industry. One effective method of monitoring biological cultures is the electrical impedance spectroscopy. In this work an integrated sensor array consisting of a four electrode sensor design on polyimide flextape in 96 well microtiter plates (MTPs) is characterised. FEM-simulations and measurements of different electrode designs are compared and show good agreement. Circular shaped electrodes are identified as optimal designs with stable signals even at high shaking frequencies. These high frequencies of at least 1000 rpm are necessary to provide high oxygen transfer in microbial cultivations. Fermentations in the MTPs were carried out with Hansenula polymorpha and Escherichia coli in filling volumes of 100–200 μL. The measured data of conductance and capacitance are shown and discussed in comparison to a reference signal obtained by a BioLector measurement system (m2p-labs GmbH, Germany). Biological phenomena, e.g. the change of carbon source of E. coli from glycerol to proteins can clearly be identified by analysing the measured capacitance and conductance during the fermentation.

[1]  J Büchs,et al.  Quasi-continuous combined scattered light and fluorescence measurements: a novel measurement technique for shaken microtiter plates. , 2005, Biotechnology and bioengineering.

[2]  E. Gheorghiu,et al.  Real-time monitoring of yeast cell division by dielectric spectroscopy. , 1999, Biophysical journal.

[3]  A. Nienow Scale-Up Considerations Based on Studies at the Bench Scale in Stirred Bioreactors , 2009 .

[4]  A. van den Berg,et al.  Monitoring of yeast cell concentration using a micromachined impedance sensor , 2005, The 13th International Conference on Solid-State Sensors, Actuators and Microsystems, 2005. Digest of Technical Papers. TRANSDUCERS '05..

[5]  J. Büchs,et al.  Effect of Oxygen Limitation and Medium Composition on Escherichia coli Fermentation in Shake‐Flask Cultures , 2004, Biotechnology progress.

[6]  Schwan Dielectric spectroscopy of biological materials and field interactions: the connection with Gerhard Schwarz , 2000, Biophysical chemistry.

[7]  E. Padan,et al.  Escherichia coli intracellular pH, membrane potential, and cell growth , 1984, Journal of bacteriology.

[8]  Jochen Büchs,et al.  Hydromechanical stress in shake flasks: Correlation for the maximum local energy dissipation rate , 2006, Biotechnology and bioengineering.

[9]  Oxygen limitation is a pitfall during screening for industrial strains , 2006, Applied Microbiology and Biotechnology.

[10]  Frank Kensy,et al.  The baffled microtiter plate: Increased oxygen transfer and improved online monitoring in small scale fermentations , 2009, Biotechnology and bioengineering.

[11]  Koji Asami,et al.  Characterization of biological cells by dielectric spectroscopy , 2002 .

[12]  J. Büchs,et al.  Characterization of gas-liquid mass transfer phenomena in microtiter plates. , 2003, Biotechnology and bioengineering.

[13]  Alvin W Nienow,et al.  The scale-up of microbial batch and fed-batch fermentation processes. , 2007, Advances in applied microbiology.

[14]  H. Schwan Electrical properties of tissues and cell suspensions: mechanisms and models , 1994, Proceedings of 16th Annual International Conference of the IEEE Engineering in Medicine and Biology Society.

[15]  H. Hashimoto,et al.  Origin of changes in electrical impedance during the growth and fermentation process of yeast in batch culture , 1989, Biotechnology and bioengineering.

[16]  J. Müller,et al.  In-situ biomass characterisation by impedance spectroscopy using a full-bridge circuit , 2001 .

[17]  Frank Kensy,et al.  High-throughput screening of Hansenula polymorpha clones in the batch compared with the controlled-release fed-batch mode on a small scale. , 2010, FEMS yeast research.

[18]  Wilfried Mokwa,et al.  Galvanic decoupled sensor for monitoring biomass concentration during fermentation processes , 2005 .

[19]  J J Heijnen,et al.  Integrated electrochemical sensor array for on-line monitoring of yeast fermentations. , 2006, Analytical chemistry.

[20]  J. Büchs,et al.  Asymmetric division of Hansenula polymorpha reflected by a drop of light scatter intensity measured in batch microtiter plate cultivations at phosphate limitation , 2009, Biotechnology and bioengineering.

[21]  Dieter Sell,et al.  Detection of the microbial activity of aerobic heterotrophic, anoxic heterotrophic and aerobic autotrophic activated sludge organisms with an electrochemical sensor , 2002, Biotechnology Letters.

[22]  Koji Asami,et al.  Characterization of heterogeneous systems by dielectric spectroscopy , 2002 .

[23]  K. Foster,et al.  RF-field interactions with biological systems: Electrical properties and biophysical mechanisms , 1980, Proceedings of the IEEE.

[24]  Urs von Stockar,et al.  On‐line biomass monitoring of CHO perfusion culture with scanning dielectric spectroscopy , 2003, Biotechnology and bioengineering.

[25]  Frank Kensy,et al.  Validation of a high-throughput fermentation system based on online monitoring of biomass and fluorescence in continuously shaken microtiter plates , 2009, Microbial cell factories.

[26]  A. E. Sippel,et al.  Development of an electronic microtiterplate for high throughput screening (HTS) , 2003, Proceedings of IEEE Sensors 2003 (IEEE Cat. No.03CH37498).