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Vol. 187, Issue 4, April 2015, pp. 113-119

 

Bullet

 

Characterization of Platinum Electrodes and In-situ Cell Confluency Measurement
Based on Current Changes of Cell-Electrodes
 

1, *Chin Fhong SOON, 2 Mee Kei FOONG, 1 Mohd Khairul AHMAD, 2 Rosliza MD ZIN, 2 Kok Tung THONG, 2 Kian Sek TEE

1 Biosensor and Bioengineering Laboratory, MiNT-SRC, Universiti Tun Hussein Onn Malaysia, 86400 Parit Raja, Johor, Malaysia
2 Faculty of Electrical and Electronic Engineering, Universiti Tun Hussein Onn Malaysia, 86400 Parit Raja, Batu Pahat, Johor, Malaysia
* Tel.: +607-4538614, fax: +607-4536060

* E-mail: soon@gmail.com

 

Received: 16 March 2015 /Accepted: 27 March 2015 /Published: 30 April 2015

Digital Sensors and Sensor Sysstems

 

Abstract: This study aimed at the development of a biosensor to examine the growth confluency of human derived keratinocytes (HaCaT) cell lines in-situ. The biosensor consists of a sputter- coated glass substrate with platinum patterns. Cells were grown on the conductive substrates and the confluency of the cells were monitored in-situ based on the conductivity changes of the substrates. Characterization of the cell proliferation and confluency were interrogated using electrical cell-substrate impedance sensing (ECIS) techniques and current change of cells using a pico-ammeter. The investigation was followed by the electrical characterization of the platinum electrode (PE) using a two probe I-V measurement system. The surface morphology of platinum electrodes were studied using an atomic force microscopy (AFM) and the HaCaT cell morphology was studied using Field-Emission Scanning Electron Microscopy (FE-SEM). The microscopy results showed that the cells coupled and proliferated on the platinum electrodes. For monitoring the conductivity and impedance changes of the cell-electrode in-situ, the cover of a Petri dish was inserted with pogo pins to be in contact with the platinum electrodes. The impedance was sampled using the ECIS technique at a twenty-four hour interval. In our findings, the cell proliferation rate can be measured by observing the changes in capacitance or impedance measured at low ac frequencies ranged from 10 - 1 kHz. In good agreement, the current measured at micro-ampere range by the biosensor decreased as the cell coverage area increased over the time. Thus, the percent of cell confluence was shown inversely proportional to the current changes.

 

Keywords: Keratinocytes, Electric Cell-Substrate Impedance Sensor (ECIS), Cell confluency.

 

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