细胞生物学：第二章细胞生物学实验技术.ppt

第二章细胞生物学实验技术;对细胞生命活动的研究成为当今生命科学发展的瓶颈;;;;;（一）普通光学显微镜;Structure of Microscope;Light Pathway of Microscope ;3. 分辨力：指分辨物体最小间隔的能力。即区分开两个质点间的最小距离。 光学显微镜的分辨力 R=0.61λ/N．A． 其中λ为入射光线波长； N．A．为镜口率 ＝ｎsinα/2； n=介质折射率； α=镜口角（样品对物镜镜口的张角） 。;表一、几种介质的折射率;显微镜的几个光学特点： 介质折射率越接近镜头玻璃的（ 1. 7 ）越好； sin α /2的最大值小于1；镜口率最大约1.6； 普通光线的波长为400~700nm，光镜分辨力约为0.2μm，人眼的分辨力为0.2mm，因此显微镜的最大有效倍数为1000X。;（二）荧光显微镜 Fluorescence microscope;Fluorescence image, DNA in blue and Microtubules in green;（三）激光共聚焦扫描显微境 Laser confocal scanning microscope, LCSM;laser confocal scanning microscope, LCSM;;LCSM Image of a Xenopus Melanophore microtubule cytoskeleton (green) and the nucleus (blue);（四）暗视野显微镜 dark field microscope;（五）相差显微镜;原理;用途：观察未经染色的玻片标本。;（六）偏光显微镜polarizing microscope;（七）微分干涉差显微镜 Differential interference contrast microscope （DIC）; ??????????????????????????????????????????????????????????????????????????????????????????Figure 3-11. Four types of light microscopy. (A) The image of a fibroblast in culture obtained by the simple transmission of light through the cell, a technique known as bright-field microscopy. The other images were obtained by techniques discussed in the text: (B) phase-contrast microscopy, (C) Nomarski differential-interference-contrast microscopy, and (D) dark-field microscopy. ;（八）倒置显微镜 inverse microscope ;（九）当代显微镜的发展趋势;二、电子显微镜;电子显微镜的基本构造;1. 原理;表二、不同光线的波长;TEM LIGHT PATHWAY;电镜与光镜的比较;电镜与光镜光路图比较;TRANSMISSION ELECTRON MICROSCOPE，TEM;2. 制样技术;2）负染技术;Examples of negtively stained and metal-shadowed specimens. Electron micrographs of a tobacco rattle virus after negtive staining with potassium phosphotungstate(a) or shadow casting with chromium(b).;3）冰冻蚀刻 freeze-etching;an onion root tip cell with no etching.;;（二）扫描电子显微镜;Figure 3-23. Scanning electron microscopy. Scanning electron micrograph of the stereocilia projecting from a hair cell in the inner ear of a bullfrog (A). For comparison, the same structure is shown by differential-interference-contrast light microscopy (B) and by thin-