Abstract
摘要 本研究主旨在於發展一個可以封裝液態活體生物樣品之元件(Wet-Cell)，使得一般電子顯微鏡可做為環境電子顯微鏡觀測生物試片。製程上我們使用微系統製程技術製作可直接用於一般電子顯微鏡樣品試片載台之元件，同時並不需要任何的改裝樣品試片載台以及外加其他機具。製程是以KOH蝕刻250um厚度的矽晶圓，並有一層100nm的氮化矽薄膜作為電子顯微鏡電子束穿透之觀測窗口。並在薄膜上鍍有一層金屬，使元件可以排除電子束轟擊下產生的電子累積，同時增強薄膜的機械強度。液態試片則以銲錫合金做為封裝材料。 捨棄過去常用之環氧樹酯做為封裝材料，而使用銲錫合金取代之，最主要因素則是金屬擁有最低的透氣率。為了使用銲錫金屬做為封裝材料，則必需在元件上鍍上一層潤濕層，增加元件與銲錫金屬的附著性。除此之外，為使銲錫合金熔化封裝，在此使用Localized Heating法，大大的降低在封裝時，元件以及液態活體生物樣品受到熱的損傷。 封裝結果可以從螢光光學顯微鏡中發現，封裝完畢後仍有活體大腸桿菌存活，但液體剩餘量大幅減少。但因目前還有許多封裝問題尚未克服，所以未來之目標則是增加封裝的成功良率，並且放進穿透式電子顯微鏡中，觀測液態活體生物試片。 Abstract The purpose of this research is to develop a Wet-Cell sealing the liquid biological sample to carry out Environmental Electron Microscopy observation. The device fabricated with MEMS technology can be used in conventional TEM holder directly, need not install any hardware additional. Fabrication is KOH etching through 250um thick silicon wafer is used to form the chamber and to release the 100nm thick Si3N4 membrane for electron beam passing through. Metal grids is used to remove the electric charge and improve the mechanical strength. Liquid sample is sealed between two cells with solder. We use solder alloy to replace epoxy as the seal material because the metal has the lower permeability. We need a wetting layer to connect Wet-Cell and solder material because we want to improve the ability of combine with tin and silicon wafer. And we use localized heating reflow solder material, to reduce the heat damage of the Wet-Cell and liquid biological sample. The result after we seal the Wet-Cell, we can find the living E.coli by fluorescence optical microscopy. But now we still have some problem did not solve, so the future goal is improve the yield, and use Transmission Electron Microscopy to observe living biological sample.