矽奈米線場效電晶體偵測活體細胞研究

Abstract

本論文探討神經分化導引晶片，藉由可重複大量製作的矽晶片微流道導引神經細胞，影響神經細胞的分化行為，並藉由觀察神經細胞培養於不同尺寸的微流道其生長分化的情形，來推論在多大的尺寸下神經細胞如何有效地被導引生長，以提供未來植入生醫晶片與現有神經組織間形成有效神經網路介面連結的解決方法。另外，本論文也運用矽奈米線場效電晶體做為高靈敏度的生物感測器，以 label-free 方式來偵測生物小分子；矽奈米線對表面電位改變有極高的靈敏度，可以作為生物感測器偵測帶電生物分子，矽奈米線會依據外圍的分子帶電荷的不同而造成其電性的改變，可以藉由觀察矽奈米線場效電晶體的電導(Conductance) 變化來作為分子的偵測。Abl tyrosine kinase 是一種磷酸轉移酶，在本實驗中將 Abl tyrosine kinase 以共價鍵方式修飾在矽奈米線的表面上，因為 Abl tyrosine kinase 對三磷酸腺苷（Adenosine triphosphate, ATP）有很好的親和力，所以可以用來偵測細胞的能量通貨 ATP 分子。最後、本實驗將 HeLa cell 培養在矽奈米線場效電晶體晶片上，直接即時偵測由自由基(ROS) 刺激後 HeLa cell 所釋放的 ATP ，證明矽奈米線場效電晶體可以即時偵測到生物體所釋放的 ATP 分子。

First part of this thesis introduces the PC-12 neural cells culture and guidance using microstructures and microfluidic channels on the silicon chip. The effectiveness of axon guidance with respect to the dimensions of microfuidic channel was explored. PC-12 neural cells were successfully cultured on microstructures and axons grew along the microfluidic channels. The results provide a pathway that bridges the organic living system and the inorganic semiconductor devices. The second part of this study utilized extremely sensitive silicon nanowire field-effect transistors(SiNW-FETs) as label-free sensors in in-situ detection of the biological small molecules, Adenosine triphosphate（ATP）. The ATP molecules were released from HeLa Cells by reactive oxygen species(ROS) stimulation. The enzyme(Tyrosine kinases, Abl) was immobilized onto the aldehyde-terminal nanowire surface; when the ATP molecules were released from the HeLa cells and binded to Abl’s binding site resulted in a conductance change of SiNW-FETs. A real-time detection of ATP molecules from ROS stimulation of HeLa cells using SiNW-FETs was also demonstrated.