利用雷射鑷夾之捕捉探針掃描顯微術

Abstract

我們結合雷射鑷夾與影像處理的技術，研發出一套能自動掃描樣品表面三維形態的顯微術。在本論文中，我們將藉由操控雷射鑷夾所捕捉的微粒子當做掃描探針，並結合即時影像重建系統，製成一套自動回授控制的三維形態掃描系統已被用來掃描一個石英玻片樣品表面的圓形圖案，此圓形圖案的直徑為6□m，我們以61 nm/sec的速率進行掃瞄，並將展示此樣品表面三維形態的影像。 另外，本論文中還介紹了兩種掃描的方法：傳統的縱向掃描以及構想中的側向掃描。我們希望結合這兩種掃描方式，能夠將樣品表面更真實的呈現。 由於雷射鑷夾的捕捉力大小在10-12牛頓的數量級，比原子力顯微鏡中大小約在10-6至10-9牛頓的凡得瓦耳作用力柔軟許多，因此，此一雷射鑷夾之捕捉探針掃描顯微術較不易傷害樣品的表面。此外雷射鑷夾原本就在水中操作，因此，雷射鑷夾掃描顯微術就比原子力顯微鏡更適合用來掃描活體生物的表面三維形態。最後，我們希望本論文所介紹的掃描式捕捉探針光學反射顯微鏡能夠應用在即時觀察活體細胞表面隨時間變化的情況。

In this thesis, we present an automatically feedback controlled trapped-probe scanning microscopy which consists of a laser tweezers system and a real-time imagine reconstruction system. In our design, we use the laser tweezers system to trap a particle as a probe. By tracking the probe with the automatic feedback control system, we can scan and reconstruct the surface of a sample. In this experiment, we scanned the circle pattern on the surface of a quartz slide, and demonstrated the reconstructed morphological image of the sample surface.

Additionally, we also introduce two kinds of scanning methods in this thesis: the traditional vertical-scanning and a novel side-scanning. We hope to combine these two scanning methods for a more precise morphological image.

Because the trapping force of optical tweezers is on the order of 1 pico-Newton (10-12 N), which is mush softer than the Van der Waals force ,10-6 N to10-9 N) of a common atomic force microscope, our optical-tweezers-based trapped-probe scanning microscope will cause less damage to the sample. Besides, operating in solution will decrease the accuracy of atomic force microscope. Therefore, this trapped-probe scanning microscope is a better tool for living cell scanning. Finally, we hope that the trapped-probe scanning microscope can be applied to observe the real time morphology of living cells.