整合介電泳細胞操控與交流電動式微幫浦之實驗室晶片應用於單細胞電穿透實驗

A Single Cell Electroporation Lab Chip via the Integration of Dielectrophoresis Manipulation and AC Electrokinetic Pump

劉邦治(Pang-Chih Liu)

指導教授 ： 劉承賢

國立清華大學/工學院/動力機械工程學系/碩士(2006年)

https://doi.org/10.6843/NTHU.2006.00521

摘要

在這篇論文研究中，我們設計一利用介電泳力 (Dielectrophoresis, DEP) 達到細胞操控並且整合交流電動式微幫浦 (AC electrokinetic pump) 之微生醫實驗室晶片，提供單一細胞電穿透 (Electroporation)的生醫應用。藉由施加不同頻率的電壓至設計的微電極上，對細胞產生正、負介電泳力，並且利用漸縮的微流道結構，達到單一細胞操控的功能，進而實現操控單細胞的電穿透功能。以交流電動式微幫浦傳送DNA或是染劑至On-Chip的電穿透實驗區域，給予交錯城堡狀電極數個脈波的電壓訊號，使得已被操控至定位的細胞其細胞膜產生暫時性的微通透孔隙，便能夠將DNA或是染劑送入細胞中，觀察其生物反應，此一技術將可提供生醫醫藥研發及癌症 (drug screening and cancer study)方面的研究應用。其研究原理，晶片設計，CFD軟體模擬與成功的實驗結果在本論文中有詳細的探討與實現。
實驗部分包含在晶片上以正、負介電泳力對細胞的操控、交流電動式微幫浦的整合並操作在特定介電泳溶液中以及單細胞的產生機制與最後電穿透生物實驗的結果都成功的實現並且整合至我們設計的生物晶片上。這份研究成功的介紹一創新的介電泳單一細胞操控機制並且整合交流電動式微幫浦應用在單細胞電穿透實驗，對於系統生物晶片的研究上提供一個創新的整合概念與應用平台。

關鍵字

介電泳 ； 交流電動式微幫浦 ； 單細胞操控

並列摘要

In this research, we report the development and experimental results of a single cell electroporation lab chip via the integration of dielectrophoresis manipulation and AC electrokinetic pump. Both dielectrophoresis force and AC electrokinetics pump are incorporated in this micro-chip for the target of single cell electroporation. By applying individual input AC voltage with different frequency on different micro electrodes, cells could be manipulated in our device based on their electric property. An additional micro-structure filter design is adopted in this chip to function as a single cell micro-channel for the purpose of centralizing the cells in a line. DNA or other nano-injectors could be pumped to the electroporation region and be inserted into the target cell via the castellated electrodes with several short electrical pulses.
The theory, chip design and simulation are reported in this thesis. The experiments including positive and negative DEP force for live cells manipulation, AC electrokinetic pump in DEP buffer, single-cell sorting mechanism, single cell electroporation and the integration of system chip functions have be demonstrated successfully. This research has demonstrated the pioneer of single-cell DEP manipulation and the integration of AC electrokinetic pump for system bio-chip research like electroporation, which could be applied to the applications of drug screening and cancer study.

並列關鍵字

Dielectrophoresis ； AC Electrokinetic Pump ； Single Cell Manipulation

參考文獻

[1] Pak Kin Wong, Tza-Huei Wang, Joanne H. Deval, and Chih-Ming Ho, “Electrokinetics in Micro Devices for Biotechnology Applications,” IEEE/ASME Transactions on Mechatronics, Vol.9, NO. 2, JUNE 2004.

[2] M. Sitti, “Survey of Nanomanipulation Systems,” Proc. of the IEEE-Nanotechnology Conference, pp.75-80, 2001

[3] G.M. Whitesides, A.D. Stroock, “Flexible methods for microfluidics,” Phys. Today 54 (6) 42–48, 2001.

[4] A. B. D. Brown, C. G. Smith, and A. R. Rennie, “Pumping of water with ac electric fields applied to asymmetric pairs of microelectrodes,” PHYSICAL REVIEW E, Vol 63, 016305, 2000.

[5] M. Mpholoa, C.G. Smith, A.B.D. Brownb, “Low voltage plug flow pumping using anisotropic electrode arrays,” Sensors and Actuators B 92 262–268, 2003.