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SEMINAR-III 28-09-09 SPEAKER KALPANA. Impedimetric monitoring of cell attachment on interdigitated microelectrodes Bin-Wha Chang a , b , Che-Hsiung Chena, Shin-Jyh Ding c , David Chan-Hen Chend, Hsien-Chang Changa , ∗

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Seminar iii 28 09 09 speaker kalpana

SEMINAR-III

28-09-09

SPEAKER

KALPANA


Impedimetric monitoring of cell attachment on interdigitated microelectrodes

Bin-Wha Changa,b, Che-Hsiung Chena, Shin-Jyh Ding c, David Chan-Hen Chend, Hsien-Chang Changa,∗

aInstitute of Biomedical Engineering, National Cheng-Kung University, Tainan 701, Taiwan, ROC

bDepartment of Healthcare Administration, Hung-Kuang University, Taichung 433, Taiwan, ROC

cInstitute of Dental Materials, Chung-Shan Medical University, Taichung 402, Taiwan, ROC

dInstitute of Veterinary Microbiology, National Chung-Hsing University, Taichung 402, Taiwan, ROC


1.INTRODUCTION microelectrodes

  • Cell attachment is the initial step in tissue or tumor formation process including cell growth, migration, junction formation, metabolism, division, differentiation, metastasis and apoptosis.

  • To assay cell attachment, direct cell counting, time-lapse cinematograph and colorimetric methods have conventionally been

    used.

    • laborious, indirect and not time-resolved.

  • Ehret et al. designed an interdigitated microelectrode to obtain the impedance data attributable to cell physiological status,the interdigitated electrode design has merits of high cell constant and uniform sensing interval.

    • very reliable and informative.


2.EXPERIMENTAL microelectrodes

Schematic diagram of the impedimetric system:


2.1. Impedimetric measuring chamber with microelectrodes microelectrodes

  • Interdigitated microelectrode plate (IMP, Fig. 1c) was patterned

    on a glass slide (26mm × 76mm; Fig. 1b) by the wet-etching process.

  • Cell chamber for impedimetric measurement was constructed by mounting and fixing a section of glass tube (12mm i.d. × 6 cm height) on the IMP plate using epoxy resin.

(b) schematic diagram of the interdigitated microelectrode plate; (c) a photograph of MDCK cells grown on the microelectrode plate.


  • RGD-C peptide was adsorbed on the microelectrodes 1 mg/cm2 by dipped-coating procedure.

    2.2. Culturing cells in the measuring chamber

  • MDCK cells (ATCC CCL-34, passages 50–90) were cultured in Dulbeco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (GIBCO) and 5% gentamycin at 37 ◦C in ahumidified atmosphere of 5% CO2 and 95% air.

    2.3. Impedance analyzing system

  • Time-resolved admittance spectra were obtained with a commercialized impedance analyzer (hp4194A).

  • The operating frequency was scanned from 1 kHz to 1MHz by 10 kHz increment, and the sampling interval was 1 min.


3. Results and discussion by dipped-coating procedure.

Monitoring of admittance during cell attachment:



The following correlation equation was established from 30 experiments

(Fig. 4) with different cell densities using chambers with bare microelectrodes:

The following equation was established using chambers with electrodes coated with RGD-C peptide (n = 20; Fig. 4):


4. Conclusions experiments

  • Data obtained with the impedimetric system were informative for cell physiology and biomaterial studies.

  • Cell attaching/detaching process, formation of confluent layer, apoptosis and even migration phenomena might be resolved and

    quantified by the proposed system.

  • The study provided also a reliable protocol to determine the cell density attached onto the surface interested and also the timing of the attach process.


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