A transparent cell-culture microchamber with a variably controlled concentration gradient generator and flow field rectifier

Cheng, Ji-Yen; Yen, Meng-Hua; Kuo, Cheng-Tzu; Young, Tai‐Horng

doi:10.1063/1.2952290

Cited by 64 publications

(33 citation statements)

References 37 publications

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“…At low flow rate of 20µL/hr, due to the low molecular weight (6045 Da) and the electrophoresis of EGF, a slight concentration difference occurred near the zigzag channel. In our previous work, we have found that the diffusion simulation is in accordance with the experimental data [48]. Therefore, cells were observed in the regions with uniform chemical concentration and EF (Figure 1).…”

Section: Methodssupporting

confidence: 85%

Evaluation of EGFR and RTK Signaling in the Electrotaxis of Lung Adenocarcinoma Cells under Direct-Current Electric Field Stimulation

et al. 2013

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Physiological electric field (EF) plays a pivotal role in tissue development and regeneration. In vitro, cells under direct-current electric field (dcEF) stimulation may demonstrate directional migration (electrotaxis) and long axis reorientation (electro-alignment). Although the biophysical models and biochemical signaling pathways behind cell electrotaxis have been investigated in numerous normal cells and cancer cells, the molecular signaling mechanisms in CL1 lung adenocarcinoma cells have not been identified. Two subclones of CL1 cells, the low invasive CL1-0 cells and the highly invasive CL 1-5 cells, were investigated in the present study. CL1-0 cells are non-electrotactic while the CL 1-5 cells are anodally electrotactic and have high expression level of epidermal growth factor receptor (EGFR), in this study, we investigated the generally accepted hypothesis of receptor tyrosine kinase (RTK) activation in the two cell lines under dcEF stimulation. Erbitux, a therapeutic drug containing an anti-EGFR monoclonal antibody, cetuximab, was used to investigate the EGFR signaling in the electrotaxis of CL 1-5 cells. To investigate RTK phosphorylation and intracellular signaling in the CL1 cells, large amount of cellular proteins were collected in an airtight dcEF stimulation device, which has advantages of large culture area, uniform EF distribution, easy operation, easy cell collection, no contamination, and no medium evaporation. Commercial antibody arrays and Western blotting were used to study the phosphorylation profiles of major proteins in CL1 cells under dcEF stimulation. We found that electrotaxis of CL 1-5 cells is serum independent and EGFR independent. Moreover, the phosphorylation of Akt and S6 ribosomal protein (rpS6) in dcEF-stimulated CL1 cells are different from that in EGF-stimulated cells. This result suggests that CL1 cells’ response to dcEF stimulation is not through EGFR-triggered pathways. The new large-scale dcEF stimulation device developed in the present work will aid the sample preparation for protein-based experiments.

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Section: Methodssupporting

confidence: 85%

Evaluation of EGFR and RTK Signaling in the Electrotaxis of Lung Adenocarcinoma Cells under Direct-Current Electric Field Stimulation

et al. 2013

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“…For example, Lin et al (2007) successfully developed an ITO based microheater chip for in situ fluid heating and thermal control in a microchip for cell immobilization application. More recently, Cheng et al (2008) proposed a similar chip with uniform temperature distribution and long-term thermal stability for cell culture purpose. The fabrication procedures of those ITO-based heating device involved the use of traditional wet etching approach and of laser-induced backside wet etching respectively to pattern ITO on a substrate.…”

Section: Introductionmentioning

confidence: 99%

Application of indium tin oxide (ITO)-based microheater chip with uniform thermal distribution for perfusion cell culture outside a cell incubator

Lin

Kuo

et al. 2010

Biomed Microdevices

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This study reports a transparent indium tin oxide (ITO)-based microheater chip and its applicability for perfusion cell culture outside a cell incubator. The attempt of the proposed ITO microheater is to take the role of conventional bulky incubator for cell culture in order to improve integratability with the experimental setup for continuous/perfusion cell culture, to facilitate microscopic observation or other online monitoring activities during cell culture, or even to provide portability of cell culture operation. In this work, numerical simulation and experimental evaluation have been conducted to justify that the presented device is capable of providing a spatially uniform thermal environment and precise temperature control with a mild deviation of +/-0.2 degrees C, which is suitable for a general cell culture practice. Besides, to testify that the thermal environment generated by the presented device is well compatible with conventional cell incubator, chondrocyte perfusion culture was carried out. Results demonstrated that the physiology of the cultured chondrocytes on the developed ITO microheater chip was consistent with that of an incubator. All these not only demonstrate the feasibility of using the presented ITO microheater as a thermal control system for cell culture outside a cell incubator but also reveal its potential for other applications in which excellent thermal control is required.

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“…[15][16][17] Both the development of CO 2 laser treatment of polymethylmethacrylate (PMMA) (Refs. 18 and 19) and the application of these modified plastics 20,21 have been reported. A recent example of the use of laser ablation for microchannel development based on a 3 layer PMMA laminate utilized two different types of lasers.…”

Section: Introductionmentioning

confidence: 99%

A parallel microfluidic channel fixture fabricated using laser ablated plastic laminates for electrochemical and chemiluminescent biodetection of DNA

Edwards

Harper²,

Polsky³

et al. 2011

Biomicrofluidics

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Herein is described the fabrication and use of a plastic multilayer 3-channel microfluidic fixture. Multilayer devices were produced by laser machining of plastic polymethylmethacrylate and polyethyleneterapthalate laminates by ablation. The fixture consisted of an array of nine individually addressable gold or gold/ITO working electrodes, and a resistive platinum heating element. Laser machining of both the fluidic pathways in the plastic laminates, and the stencil masks used for thermal evaporation to form electrode regions on the plastic laminates, enabled rapid and inexpensive implementation of design changes. Electrochemiluminescence reactions in the fixture were achieved and monitored through ITO electrodes. Electroaddressable aryl diazonium chemistry was employed to selectively pattern gold electrodes for electrochemical multianalyte DNA detection from double stranded DNA (dsDNA) samples. Electrochemical detection of dsDNA was achieved by melting of dsDNA molecules in solution with the integrated heater, allowing detection of DNA sequences specific to breast and colorectal cancers with a non-specific binding control. Following detection, the array surface could be renewed via high temperature (95 C) stripping using the integrated heating element. This versatile and simple method for prototyping devices shows potential for further development of highly integrated, multi-functional bioanalytical devices.

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A transparent cell-culture microchamber with a variably controlled concentration gradient generator and flow field rectifier

Cited by 64 publications

References 37 publications

Evaluation of EGFR and RTK Signaling in the Electrotaxis of Lung Adenocarcinoma Cells under Direct-Current Electric Field Stimulation

Evaluation of EGFR and RTK Signaling in the Electrotaxis of Lung Adenocarcinoma Cells under Direct-Current Electric Field Stimulation

Application of indium tin oxide (ITO)-based microheater chip with uniform thermal distribution for perfusion cell culture outside a cell incubator

A parallel microfluidic channel fixture fabricated using laser ablated plastic laminates for electrochemical and chemiluminescent biodetection of DNA

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