本研究以VLSI技術在矽晶片上製備出線型以及洞型有序陣列，再利用聚二甲基矽氧烷(Polydimethylsiloxane，PDMS)將矽晶片上的圖案轉印出來成為PDMS的一維以及二維光柵。接著將表面進行親水處理、矽烷類自組裝層改質、以EDC/NHS反應將上皮細胞黏附分子抗體-生物素(Anti-Epithelial Cellular Adhecion Molecule-Biotin，Anti-EpCAM-biotin)中biotin之羧基活化固定於PDMS光柵上，用以捕捉血液中循環腫瘤細胞(Circulating tumor cells，CTCs)，由於血液分離術中無法分離CTC與白血球，因此搭配牛血清蛋白(Bovine Serum Albumin，BSA) 處理降低白血球沾黏；光柵的結構可雷射產生特殊的繞射現象，可以繞射強度為基準進行大腸癌快速檢測。結果顯示，基材對白血球的沾附量可達到一穩定值(1000萬顆/c.c.血液的濃度維持在平均178顆)，雷射分析儀得到健康人繞射強度值範圍(3.33173 E+07 cnts ~3.45794 E+07 cnts)；另外繞射強度值偵測靈敏度為4顆，且隨著細胞培養顆數的上升，繞射強度呈下降的趨勢；在臨床試驗中，可以經由血液快速分辨健康人及大腸癌患者。
臨床檢體之癌細胞有利於未來基因治療的研究，故藉由細胞培養放大癌細胞數量就變得非常重要，且因為一般細胞生長監控用計數器極為不便，需要耗費人力及時間且非常主觀，所以以雷射分析的方式取而代之。為了將基材的培養效能提升，選用細胞外基質之高分子作聚電解質多層膜的材料，增加細胞的增生及貼附率，並搭配雷射分析儀作細胞生長監控，結果顯示細胞外基質環境確實提升細胞的貼覆效果，分別從60.4%(柱型)、71%(線型)上升到70.7%(柱型)及92.3%(線型)。雷射數值對應細胞顆數成長也呈高度線性關係，證實雷射分析儀可以進行細胞監控定量。依照細胞成長趨勢到第十天變化量為十倍，推估當循環腫瘤細胞抓到10顆後，在第十天可以成長到100顆。最後以多層膜之裂解酶對培養後之細胞作釋放，釋放率達到98.9%(線型)及99.7%(柱型)，成為一可偵測又能培養的釋放之基材。

In this study, we patterned ordered line and hole arrays with photoresist on the silicon surface by VLSI process. The regular patterns of photoresist were transferred with polydimethylsiloxane (PDMS) to generate ordered line and pillar arrays of 1D and 2D grating of PDMS. The grating surfaces were sequentially modified with Anti-Epithelial Cellular Adhecion Molecule-Biotin(Anti-EpCAM-biotin), a kind of antibody for immunosorbent of CTC, by EDC/NHS reaction. In addition, bovine serum albumin (BSA) was exploited to educe the fouling of white blood cells(WBCs). Regular structure leads to special diffraction effect, CTC attachment results the decrease of the diffraction intensity. Therefore, a laser system can exploit to detect the CTCs rapidly on the surface. The results showed that the amounts of WBCs attached on substrate remained 178corresponding to the diffractive intensity ranged from 3.33173 E+07 cnts to 3.45794 E+07 cnts for the sample from a healthy subject. The diffractive intensity decreased with the attachment of CTCs on the surface. The sensitivity of the CTC detection reached to 4 cells. The results of clinical experiment suggest that the detection of CTCs with the approach is able to diagnose a healthy subject and a patient of colorectal cancer.
The cancer cells from clinical specimens have a great development of future genetic therapy, cell culture becomes extremely important. In general, monitoring of cell growth was inefficient by using cell counter; however, it is inconvenient. Therefore, laser system was exploited to monitor the cell amounts in real time. In order to increase the efficiency of cell culture, we used layer by layer to coat extracellular matrix polymers on the substrate. The laser value corresponds to the numbers of cells reached a highly linear relationship, which could quantify cells. According to the trends of cell proliferate rate, we estimated that 10 CTCs captured from clinical specimens may increase up to 100 cells in 10 days. Finally, we released the cells, and 98.9% (line array) and 99.7% (pillar array) cells on the surface could be released.
The 1D and 2D grating of PDMS can capture the CTCs from the whole blood samples and culture the clinical CTCs, which have potential applications as diagnosis of colorectal cancer.

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