本實驗以1.01μm聚苯乙烯球作為奈米遮罩，利用奈米球自組裝微影術成功製備出高整齊度且規則排列的二維奈米金球陣列。在製備聚苯乙烯球自我組裝遮罩部分，本實驗嘗試兩類方法：(1)方便大量製備的旋佈法；及(2)可控制晶格成長方向的微圓洞液滴蒸發自組裝法。在旋佈法部分，藉由添加酒精溶劑或基板氧電漿處理兩種分法來增加聚苯乙烯球單分子層排列面積。實驗發現添加酒精會使溶劑揮發增快，粒子傾向多晶排列，最大單晶晶格僅約5μm*5μm。在氧電漿旋佈法部分，探討旋佈轉速、濃度及粒徑均勻度對遮罩製備影響。實驗發現當旋佈轉速下降或粒子濃度提升時，皆可增加聚苯乙烯球排列面積，在實驗最佳參數下，單分子層幾乎可全部均勻覆蓋1.5cm*1.5cm之玻璃基板上。此外，粒子均勻度也是製備高品質遮罩之關鍵，使用粒子均勻度高之懸浮液做旋佈，則可得到最大單晶晶格約為50μm*50μm。在微圓洞液滴蒸發自組裝法部分，其機制是液滴在PDMS微槽蒸發時，會從原本凸起的半圓形轉變為下凹的新月形，因為這個特性使聚苯乙烯球可從中央開始成核，隨著液體慢慢蒸發，再逐漸向外成長晶格，因為其晶格成長具有方向性，因此能製備出晶格缺陷更少，單晶區域更大的聚苯乙烯球遮罩，藉由調控濃度與側壁高度，可製備出250μm*150μm之單晶晶格。完成遮罩後，以熱蒸鍍方式沉積10 nm金薄膜，經過850度高溫回火後，可得到平均粒徑135nm之金球陣列，若改變蒸鍍角，則可以成功製備出不同尺寸之六角形規則排列奈米金球陣列。實驗最後利用硫醇分子將螢光蛋白質修飾奈米金球陣列，使用100倍油鏡螢光顯微鏡可清楚觀測到六角形排列之螢光影像，證實蛋白質分子有成功接枝至金球表面，初步完成奈米金陣列生物檢測晶片之製備。

This experiment applied nanosphere lithography (NSL) to fabricate periodic nanogold arrays by using self-assembly two-dimensional colloidal crystals as the lithographic masks. We have tried two methods to self-assemble1.01μm polystyrene latex colloids. One is spin-coating, which is convenient and inexpensive. Another is “droplet evaporation self-assembling in microwells” , which could control the direction of crystals growth by the meniscus formed during the evaporation process. In the spin-coating part, we increased the monolayer areas by adding solvent alcohol or treating the substrate with O2 plasma before spinning. The biggest single crystal domain we observed in adding alcohol method was only 5μm*5μm, while in O2 plasma treating method was 50μm*50μm. In the droplet evaporation part, the biggest single crystal domain we observed was 250μm*150μm.
After finishing the preparation of masks, we deposited 10 nm gold thin films onto the masks by thermal evaporation. Removing the masks and annealing the samples 2 hours in 850℃, we could obtain the densely packed two-dimensional ordered nanoarrays of gold nanospheres. Then we adjusted the angle of thermal evaporation to fabricate different sizes of uniform gold nanospheres arrays from average diameters 135nm to 72 nm. In the last part, we have tried to conjugate the fluorescent proteins on the gold nanostructures and have successively observed the signals by using 100X optical microscopy.

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