RNA之完整性在生物領域中扮演著很重要的角色，因為純度不佳之RNA可能對下游實驗結果造成很大的影響。在此研究中發展了一種快速及經濟之方法分析總RNA（total RNA）之完整性，是以毛細管電泳暨雷射誘發螢光在電滲流存在下（capillary electrophoresis with laser-induced fluorescence in the presence of electroosmotic flow）為基礎進行分析。此方法利用PEO（Mave 4,000,000 Da）作為分離RNA之篩分介質，並將溴化乙錠（微克/毫升）溶解在聚合物溶液作為標定RNA之染料。成功地在25分鐘內完全分離出總RNA中的 28S、18S、5.8S、5S核醣體RNA（robosome RNA）及傳遞RNA（transfer RNA）。此研究中，利用線上濃縮技術注入大體積樣品可增加實驗靈敏度，使實驗可偵測到最低28S及18S rRNA濃度為100 pg/μl。最後利用加熱不同時間成功監測RNA的降解情況。結果顯示，使用毛細管電泳暨雷射誘發螢光在電滲流存在下可有效的分析RNA之完整性。實驗的第二部分是組裝暗視野顯微鏡偵測金奈米粒子。利用高數值接物鏡（high numerical objective）、擋光片（field stop）以及視野限制（iris），成功地觀察32 nm金奈米粒子的布朗運動（brownian motion），並利用商業化相機收集此影像。結果顯示，此儀器可以用來偵測以金奈米為基礎的奈米感測器（nanosensing）。 RNA integrity plays an important role in RNA studies because poor RNA quality may have a great impact on downstream methodologies. This study proposed a rapid, and cost-effective method to analyze RNA integrity based on capillary electrophoresis with laser-induced fluorescence(CE-LIF)in the presence of electroosmotic flow. This proposed method used the poly(ethylene) oxide (Mave 4,000,000 Da) as sieving matrix for total RNA separation. The ethidium bromide (μg/mL) was dissolved in polymer solution as interchelating dye for the on column fluorescent labeling. The separation of the 28S rRNA, 18S rRNA, 5.8S rRNA, 5S rRNAs and tRNA from cell extracted human total RNA was accomplished within 25 minutes. In this work, online concentration technique was used to increase the sensitivity by larger volume sample injection. The lower detectable concentration of 28S and 18S rRNAs is 100 pg/μL. In addition, the temperature-induced degradation of total RNAs could be monitored by CE-LIF. The result demonstrated the separation of total RNA by CE-LIF in the presence of EOF is useful for determination of RNA quality. The second part of this thesis is to build a dark field microscope to detect gold nanoparticles. By using high numerical objective, appropriately field stop and iris, observation of brownian motion of 32 nm gold nanoparticles was successfully recorded by commercial CCD camera. This result indicated the proposed instrument may potentially be use for sensitive detection of nanoparticle-based nanosensing.
