本研究利用半導體加工製程技術製備微電極陣列感測探針，製程的部分共分成為三個階段，第一階段為金屬圖案之定義，使金屬導線，封裝端與電極端的圖案成形。第二階段為探針導線區域絕緣，對整片晶圓沉積介電層後，運用蝕刻技術將電極端與封裝端之金屬裸露出來，僅剩金屬導線絕緣達到部分區域絕緣之目的。第三階段為探針邊界輪廓定義，運用微影製程將探針邊界定義出來，再搭配蝕刻製程將探針從矽晶圓上取下。成功取下之再探針經過封裝、表面修飾與酵素塗佈後製備成榖氨酸感測探針，並透過一系列的效能測試，包括干擾物測試、線性範圍測試、穩定度測試以及靈敏度測試等，評估所製備的榖氨酸感測探針是否可進行植入型動物實驗。最後將榖氨酸感測探針植入白鼠腦部丘腦的區域，對白鼠後肢與尾巴進行痛覺刺激，藉由監控丘腦瞬時的榖氨酸釋放量，來達到研究神經傳遞途徑的目的。本研究不僅成功製備出微電極陣列感測探針，並在動物實驗上得到榖氨酸感測訊號與實驗結果，證明神經傳導途徑為對側傳導。由於植入型動物實驗的成功，因此可預期未來是可以應用在許多神經傳導物質研究的領域。

In this study, we used the semiconductor manufacturing technology to prepare microelectrode array (MEA) probes which were used for the fabrication of glutamate biosensors and the application for in vivo tests. The semiconductor manufacturing process to prepare MEA probes was divided into three parts. The first part was formation of the specific pattern of platinum metal layer on the silicon wafer. This specific pattern was to define the body of MEA probes which includes electrode sites, channels and bonding pads. The second part was to insulate the channels. We used plasma enhanced chemical vapor deposition (PECVD) to deposit the dielectric layer (polypyrrole and Nafion®) on the wafer, then we used the etching process to etch electrode sites and bonding sites so there were insulation layers on the channels The third part was to define the outline of MEA probes. We used photolithography process to define the outline region, and then we uses inductively coupled plasma (ICP) to etch the outline. Deep etching through the wafer was required so that MEA probes could be taken off from the wafer. After we finish the process for the preparation of MEA probes, it was required to modify the electrode surface with permselective polymer layers and the glutamate oxidase layer. The glutamate oxidase layer can make the glutamate sensor with better specificity and permselective layers can prevent signals form interferent.
The goal of this study is to apply this glutamate biosensor in vivo for real-time monitoring the concentration changesof glutamate, and correlate the glutamate release in thalamus of rates to the noxious stimuli through spinothalamic tract. Before in vivo test, it was required to evaluate the performance of glutamate biosensors that we fabricated by several tests to confirmation the feasibility of its application in vivo (experimental results was shown in Chapter 4). Finally, the glutamate biosensor was implanted in the thalamus of rat’s brain for monitoring the release of glutamate before and after the noxious stimuli. Calibration cures of the glutamate sensor before and after the implantation were established (experimental results was shown in Chapter 5).
In this study, we demonstrated a sensitive, small, and fast glutamate sensor based on the MEA probe fabricated by semiconductor manufacturing. The in vivo study confirmed that the ascending pathway of spinothalamic tract was contralateral.

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