氮化矽微米孔隙元件陣列之癌細胞捕獲與量測研究

Abstract

本研究利用氮化矽微米孔洞陣列作為細胞濃度過濾與偵測之元件。首先，利用微米製作技術製備出氮化矽微米孔洞陣列，孔洞數分別為(矩形30x15)、(圓形31x31)、(圓形10x10)和(圓形7x7)。經由微流道模擬的建立分析以及實際過濾及量測結果。實驗過程中，檢體分別以固定流速，50和37 µl min-1 的流速進入流道中過濾及量測，電壓由鎖相放大器經由氯化銀參考電極供給並且同時量測電流值變化，操作條件為10 mV，頻率10 Hz。實驗過程中，以不同的元件過濾和量測檢體中粒子濃度，並且比較其結果。結果顯示，擁有(圓形7x7)的元件可到達最低濃度的偵測極限，其對於聚苯乙烯粒子以及子宮頸癌細胞的偵測最低濃度約為100 particles/ml 。此外，本研究成功的結合可以快速過濾與量測粒子濃度的系統。

In this thesis, silicon nitride micropore array was applied for resolving bead and cell concentrations by filtering and measuring for large volume analytes. First, the micropore array on free standing silicon nitride membrane was fabricated via microfabrication processes with four different pore numbers and shapes, rectangle(30x15) and circle(31x31,10x10,7x7). Microfluidic COMSOL simulation of the micropore array device was constructed and compared with the filtration and measurement data. Samples were flowed through the micropores at a flow rate of 50 and 37 µl min-1 via a syringe pump. The AC sine-wave voltage (10 mV rms) at 10 Hz was applied between two Ag/AgCl electrodes across the micropores by a lock-in amplifier, and the ionic current was analyzed. Devices with different pore numbers and shapes were optimized, and applied to resolve the beads/HeLa cells concentrations. The results showed that the device with micropores (circle7x7) exhibited the best detection limit. Concentratons of polystyrene beads and HeLa cells down to 100 particles/ml were demonstrated.