Fluorescence Recovery After Photobleaching, FRAP 荧光漂白恢复原理:利用高能量激光束的照射使特定的区域的荧光发生不可逆 的淬灭,通过非漂白区的荧光标记分子在膜上或胞浆中运动至光漂白区来完成 光漂白区荧光的恢复。FRAP可以用于解决活细胞内包括蛋白定位,动力 学以及与其他成分相互作用等一系列问题。 初始状态 光漂白 恢复 F Fc
Fluorescence Recovery After Photobleaching，FRAP 荧光漂白恢复原理: 利用高能量激光束的照射使特定的区域的荧光发生不可逆 的淬灭，通过非漂白区的荧光标记分子在膜上或胞浆中运动至光漂白区来完成 光漂白区荧光的恢复。FRAP 可以用于解决活细胞内包括蛋白定位，动力 学以及与其他成分相互作用等一系列问题
IVideo-enhance(contrast) microscopy E (b) Figure A-17 Computer-Enhanced Digital Video Microscopy image resulting from electronic contrast enhancement of the original images obtained with light microscopy. In this example, an image of image in (a), which is then (c)subtracted from image (a, leaving only the several microtubules, which are too small to be seen with unenhanced microtubules. (d) The final, detailed image resulting from electronic light microscopy are processed to make them visible in detail. (la)The averaging of the separate images processed as shown in a-c Observing living specimens Greatly increase the contrast of an image so that very small objects become visible
Video-enhance(contrast) microscopy ❖Observing living specimens; ❖Greatly increase the contrast of an image so that very small objects become visible
2. Transmission Electron Microscopy A. The comparison of the lens systems of LM and tem 电子显微镜与光学显微镜的基本区别 分辨本领 光源 透镜 真空 成像原理 光学显微 200nm 可见光 玻璃透镜不要求真空利用样本对光的吸收 镜 (波长400 形成明暗反差和颜色 700nm) 变化 电子显微 接近0.1nm电子束 电磁透镜1.33×103~利用样品对电子的散 镜 波长0.01 1.33×105Pa射和透射形成明暗反 0.9nm) The resolving power for LM; The resolving ability(0. 2nm) and resolving power(5nm) for TEM
分辨本领 光 源 透 镜 真 空 成像原理 光学显微 镜 200nm 可见光 (波长400～ 700nm) 玻璃透镜 不要求真空 利用样本对光的吸收 形成明暗反差和颜色 变化 电子显微 镜 接近0.1nm 电子束 (波长0.01～ 0.9nm) 电磁透镜 1.33×10-3～ 1.33×10-5Pa 利用样品对电子的散 射和透射形成明暗反 差 电子显微镜与光学显微镜的基本区别 The resolving power for LM; The resolving ability(0.2nm) and resolving power(5nm) for TEM 2. Transmission Electron Microscopy A. The comparison of the lens systems of LM and TEM
息子显微镜 阴极(灯丝) 电子枪 阳极 聚光镜 聚光镜 样品 样品室 物镜 中间像 中间镜 投影镜 投影镜 观察窗 荧光屏 荧光屏成像 电子显微镜光成像原理 A.透射电子显微镜剖面图。B.透射电子显微镜电子成像原理图
电子显微镜光成像原理 A.透射电子显微镜剖面图。B. 透射电子显微镜电子成像原理图
B Specimen Preparation for Electron Microscopy ☆ Thin Sectioning for TEM dehydrated by placing solution. centrations acetone or alcchol The wax sections: 5um The Plastic ultrathin sections for TEM: 50- when the plastic is hard. the 100nm ready for sectioning. Tissue is placed in final astc is polymerized Sections of lM:>5um he knie onto the surface of a water trough Sections of TEM: <0.1 u ms sections are picked
B. Specimen Preparation for Electron Microscopy ❖Thin Sectioning for TEM The wax sections: 5um; The Plastic ultrathinsections for TEM: 50- 100nm Sections of LM: >5um; Sections of TEM: <0.1 μ ms