Development of Transparent Bio-Microfluidic Chip for Study of Cell Behavior Under Microcontrolled Environment
Date Issued
2010
Date
2010
Author(s)
Yen, Meng-Hua
Abstract
Experiments with transparent bio-microfluidic chip can provide essential real-time observation for cell growth and cell studies. Therefore, this study developed a transparent bio-microfluidic chip for study of cell behavior under microcontrolled environment. This study developed transparent microfluidic chips by laser direct-write methods (LDW), which are the laser-induced backside wet etching (LIBWE) systems and CO2 laser machining. Then, this study successfully used these methods to develop a transparent bio-microfluidic chip with chemotaxis or electrotaxis for study of cell behavior. In addition, this study develops two patterning techniques, ITO patterning and cell patterning, to apply in study of cell behaviors.
Transparent microfluidic chips include glass and PMMA microfluidic chip. For glass microfluidic chip, four fabrication methods are used for fabricating. These methods, which include modified CO2 laser machining, UV LIBWE using organic absorber, visible LIBWE using organic absorbers and visible LIBWE using metallic absorbers, can produce crack-free and high-quality microchannels. For PMMA microfluidic chip, visible LIBWE using the metallic absorber is used to fabricate and also produces high-quality microchannels. These four fabrication methods are LDW methods, so the overall development time of glass or PMMA microfluidic chip is shortened to less than 24 h. In addition, the mechanism of LIBWE is discussed in depth because the LIBWE in this dissertation is an important fabrication method.
This study presents an autonomous microfluidic chip for long-term and real-time observation of cell migration by a microscope. It was combined the microchannels and ITO electrodes to form a closed microchamber with chemical gradient for cell culturing and observation. For chemotaxis collaborated with the Miss Hsu, we used a super-resolution microscopy technique, non-interferometric wide-field optical profilometry (NIWOP), to observe filopodium activity of cells under the EGF gradient. Higher filopodium activity is observed at the side facing higher EGF concentration for single CL1-0 cell subjecting to the EGF gradient. For electrotaxis collaborated with the Miss Huang, we enabled observing the electrotactic response of lung cancer cells by a sealed culture chamber that is suitable for long-term electrotaxis study with a microscope. We used lung cancer cell lines with high and low metastasis potential, CL1–5 and CL1–0, respectively, to demonstrate the function of the electrotactic chip.
Finally, this study developed the ITO patterning by visible LIBWE and the cell patterning by CO2 or UV laser machining. The obtained ablation result from ITO patterning by visible LIBWE excels that from front-side laser ablation. The ITO patterning was then utilized in fabricating a transparent gas flow meter. The cell patterning was a simple and effective method for patterning cells on a glass substrate. A passivation layer that is capable of preventing cell adhesion was first coated onto glass surface and then cells adhere and grow cleanly in the laser defined pattern.
Subjects
Laser-induced backside wet etching(LIBWE)
CO2 laser machining
ITO patterning
cell patterning
cell culture system
electrotaxis
chemotaxis
SDGs
Type
thesis
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