Microfluidic Device for Rapid Medium Exchange for Cellular Electrophysiology Study
Date Issued
2006
Date
2006
Author(s)
Liu, Kuan-Ting
DOI
en-US
Abstract
The integrated patch clamp chip consisting of microfluidic system is a powerful tool for the study of ion channel. The goal of this thesis is to develop a rapid medium exchange platform for electrophysiology study to achieve high-throughput of ion channel signals. The platform consists of micro-mixer and planar patch clamp chip. Thermal reflow of glass is applied to generate very smooth surface on the chip by utilizing the CO2 laser to penetrate the borosilicate glass. A special discovery of aperture is the hourglass shaped aperture. Immobilized cells fit perfectly to the hourglass apertures offering a large contact area for sealing between the cell membrane and the surface of aperture. Borosilicate glass was chosen due to that it is inexpensive and good electrical property. Micro-mixer is made of polydimethylsiloxane (PDMS) by using soft lithography technology. Different mediums can be obtained by controlling the injected flow rate of syringe pump and then flowed through the trapped cell. Verification of the device used blue food dye and clean water as working fluid at 16mm/s.
Results show that the rapid medium exchange platform can provide different concentration from 0% to 100% with increment of 25%. Exchanging times from 0~25% is almost 25seconds and other steps are almost 5seconds individually. Seal resistance of Chinese hamster ovary (CHO) achieved 200~300Mohm steadily. Flow that can pass through the cell membrane cannot affect the seal resistance of the trapped cell with any other flow rate of syringe pump. However, accuracy about fabrication of aperture and the yield of giga seal are not as good as expectation. The stability of laser engraver, cell preparation and the parameters of seal test are believed to be the key problems that need to be improved. The microfluidic device for rapid medium exchange can succeed to achieve high-throughput for electrophysiology.
Subjects
電生理
微流體
溶液交換
雷射鑽孔
濃度產生器
漏斗狀孔洞
electrophysiology
patch clamp
microfluidic
medium exchange
laser drilling
concentration generator
hourglass-shaped aperture
Type
thesis
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