Low-cost microelectrode array with integrated heater for extracellular recording of cardiomyocyte cultures using commercial flexible printed circuit technology

Abstract This article reports the use of commercial, flexible printed circuit technology for the fabrication of low-cost microelectrode arrays (MEAs) for recording extracellular electrical signals from cardiomyocyte cultures. A 36-electrode array has been designed and manufactured using standard, two-layer, polyimide-based flexible circuit technology, with electrode diameters of 75 and 100 μm. Copper structures defined on the backside of the array have been used for low-power thermal regulation of the culture. Electrical characterization of the gold-plated electrodes showed impedances below 250 kΩ at 1 kHz. Functional testing was conducted using HL-1 cardiac myocytes. The arrays proved biocompatible, and supported the formation of functional syncytia, as demonstrated by electrical recordings of depolarization waves across the array. A comparison with conventional, glass-based MEAs is presented, which reveals differences in signal strength (smaller for larger electrode) and variability (less for larger electrodes), but no effect of the substrate types on culture parameters such as beat rate or conduction velocity. The performance of the on-chip heating was evaluated, with typical temperature settling times (to ±0.1 °C) below 10 s, for a power consumption around 1 W (at 37 °C). Accuracy and stability are discussed. HL-1 cell responses to various temperature profiles enabled by the on-chip heating are presented, showing a remarkable correlation between temperature and beat rate.

[1]  A. Curtis,et al.  Effective extra-cellular recording from vertebrate neurons in culture using a new type of micro-electrode array , 2002, Journal of Neuroscience Methods.

[2]  Analysis of Microelectrode-Recorded Signals from a Cardiac Cell Line as a Tool for Pharmaceutical Screening , 2001 .

[3]  R. S. Pickard,et al.  Flexible printed-circuit probe for electrophysiology , 1979, Medical and Biological Engineering and Computing.

[4]  U. Egert,et al.  A thin film microelectrode array for monitoring extracellular neuronal activity in vitro. , 1994, Biosensors & bioelectronics.

[5]  E. Perl,et al.  Microelectrode arrays for stimulation of neural slice preparations , 1997, Journal of Neuroscience Methods.

[6]  Ulrich Egert,et al.  Biological application of microelectrode arrays in drug discovery and basic research , 2003, Analytical and bioanalytical chemistry.

[7]  N J Izzo,et al.  HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Guenter W. Gross,et al.  Recording of spontaneous activity with photoetched microelectrode surfaces from mouse spinal neurons in culture , 1982, Journal of Neuroscience Methods.

[9]  Gregory T. A. Kovacs,et al.  Electronic sensors with living cellular components , 2003, Proc. IEEE.

[10]  D. Robinson,et al.  The electrical properties of metal microelectrodes , 1968 .

[11]  Xiangqin Cui,et al.  Sensors and Actuators B , 2003 .

[12]  H. Oka,et al.  A new planar multielectrode array for extracellular recording: application to hippocampal acute slice , 1999, Journal of Neuroscience Methods.

[13]  Laurent Giovangrandi,et al.  Sensitivity of cell-based biosensors to environmental variables. , 2005, Biosensors & bioelectronics.

[14]  K H Gilchrist,et al.  General purpose, field-portable cell-based biosensor platform. , 2001, Biosensors & bioelectronics.

[15]  B D DeBusschere,et al.  Portable cell-based biosensor system using integrated CMOS cell-cartridges. , 2001, Biosensors & bioelectronics.

[16]  C. Chow,et al.  Copper toxicity, oxidative stress, and antioxidant nutrients. , 2003, Toxicology.

[17]  Robert A. Malkin,et al.  A simulation study evaluating the performance of high-density electrode arrays on myocardial tissue , 2000, IEEE Transactions on Biomedical Engineering.

[18]  R A Malkin,et al.  Construction of a very high-density extracellular electrode array. , 2000, American journal of physiology. Heart and circulatory physiology.

[19]  B. Wheeler,et al.  A flexible perforated microelectrode array for extended neural recordings , 1992, IEEE Transactions on Biomedical Engineering.

[20]  G. G. Stokes "J." , 1890, The New Yale Book of Quotations.

[21]  Gregory T. A. Kovacs,et al.  IMPEDANCE IMAGING FOR HYBRID BIOSENSOR APPLICATIONS , 1996 .

[22]  R. Ideker,et al.  Estimation of conduction velocity vector fields from epicardial mapping data , 1998, IEEE Transactions on Biomedical Engineering.

[23]  Marc Olivier Heuschkel Fabrication of multi-electrode array devices for electrophysiological monitoring of in-vitro cell/tissue cultures , 2001 .