光學顯微技術
Advanced light microscopy
| 節 | 週四 |
|---|---|
3 10:10–11:00 | 光學顯微技術 YL837 2 節連堂 |
4 11:10–12:00 |
* 根據陽明交大上課時間表所列
光學顯微鏡從十六世紀發明以來,已經過了幾百年的歷史,近年來,由於各式光學技術、分子標記技術、以及數位顯像技術的演進,讓光學顯微鏡從靜態的組織染色切片,發展到動態的細胞、組織甚至是活體生物的觀察,並從利用染色對結構強度對比的偵測,發展到對光波相位、光譜、非線性光學現象的偵測。近年來更利用先進的單分子標記技術、非線性光學、及近場光學試圖突破光學繞射限制,來進行超解像力顯微造影。本課程第一部份將從顯微鏡發明的歷史、顯微鏡基本構造、常用顯微鏡對比方法,配合淺顯易懂的理論,讓同學對顯微鏡使用及原理友更進一步的認識,幫助同學拍出更好的高品質影像,讓顯微鏡使用不再成為夢靨。課程第二部份將介紹最新的光學顯微鏡技術,包括共軛焦顯微鏡、多光子顯微鏡、FRET (fluorescence Resonance Energy Transfer)、FLIM (Fluorescence Lifetime Imaging Microscopy)、超解像力光學顯微鏡、光學顯微操控等最先進之科技,及其在生物醫學研究上的應用。本課程也會在課程當中穿插顯微鏡實際操作及演示,讓同學有機會接觸顯微鏡,並將其功能發揮到淋漓盡致。Light microscopy has been evolved for several hundreds years and has tremendous impacts on biology, medical research and other fields in science. In the past few decades, light microscopy has advanced from observing the structures of microorganisms, cells, and traditional histological tissue slices to dynamically recording behaviors of microscopic structures in live cells, tissue, or even organisms, thanks to the development of novel optical instrumentation, laser, non-linear optics, near-field optics, and new labeling technologies. Recently, super resolution microscopy has also been built to overcome the optical limitation in resolution. The first part of the course is designed to give students an introduction of the principles behind conventional microscopy. The second part will cover recent development of new technologies in microscopy, including confocal microscopy, multi-photon microscopy, fluorescence resonance energy transfer (FRET), fluorescence lifetime imaging microscopy (FLIM), total internal reflection fluorescent (TIRF) microscopy, and techniques in super-resolution microscopy. The course will also include lab demonstrations for the students to have hands-on experience to master advanced microscopy techniques.
期中考 30%,期末報告30%,期末考30%,課堂表現10% Midterm Exam 30%, Oral Presentation 30%, Final Exam 30%, Attendance 10%
| 週次 | 主題 |
|---|---|
| 第 1 週 | 緒論: 光學與成像原理Introduction: Optics and Imaging |
| 第 2 週 | 光學顯微鏡基本構造Basic components of a light microscope |
| 第 3 週 | 常用光學顯微鏡技術: Bright field, Dark field, Rheinberg illumination, Phase contrastTechniques in microscopy: Bright field, Dark field, Rheinberg illumination, Phase contrast |
| 第 4 週 | DIC顯微鏡與實習Differential interference contrast (DIC) microscopy |
| 第 5 週 | 螢光顯微鏡與實習Fluorescence microscopy and lab |
| 第 6 週 | 共軛焦顯微鏡與實習Confocal microscopy and lab |
| 第 7 週 | 螢光蛋白與分子螢光探針之發展Development of fluorescent proteins and probes |
| 第 8 週 | 多光子顯微鏡與實習Multi-photon microscopy and lab |
| 第 9 週 | 期中考Midterm exam |
| 第 10 週 | 高等螢光顯微鏡:Spectral imaging, FRAP, FLIP, and FSMSpectral imaging, FRAP, FLIP, and FSM |
| 第 11 週 | 活細胞顯微系統與Deconvolution microscopyLive cell imaging and deconvolution microscopy |
| 第 12 週 | 特殊光學顯微鏡:FRET, FLIMFRET, FLIM, and Optical tweezers |
| 第 13 週 | 特殊光學顯微鏡:Optical tweezers, Total internal reflection fluorescence (TIRF) microscopyTotal internal reflection fluorescence (TIRF) microscopy, light-sheet microscopy, and Fluorescence correlation spectroscopy (FCS) |
| 第 14 週 | 特殊光學顯微鏡:light-sheet microscopy, and Fluorescence correlation spectroscopy (FCS)Superresolution Microscopy: Near-field scanning optical microscope (NSOM), Stochastic optical reconstruction microscopy (STORM), Photoactivated localization microscopy (PALM), Stimulated emission depletion (STED) microscopy |
| 第 15 週 | 超解像力顯微鏡:Near-field scanning optical microscope (NSOM), Stochastic optical reconstruction microscopy (STORM), Photoactivated localization microscopy (PALM), Stimulated emission depletion (STED) microscopyStructured illumination microscopy (SIM), Super-resolution Optical Fluctuation Imaging (SOFI) |
| 第 16 週 | 期末考 Final exam |
| 第 17 週 | 彈性上課 |
| 第 18 週 | 彈性上課 |