冷冻电镜.ppt

1、制作 :丁宇航组员：谭志洪 张康 吴京洋 苟雪莲 刘雄 朱瑜我们或许在不久的将来就能获得生命复杂机制的原子级分辨率的图片了。这正是 2017 年的诺贝尔化学奖，授予 Jacques Dubochet, Joachim Frank 和 Richard Henderson，因他们在冷冻电子显微镜方面的卓越贡献，他们将冷冻电子显微镜技术简化，并将其应用在生物分子成像方向。此种成像技术将生物化学领域推进了新时代。Contents目录The History of Application of cryo-Electron Microscope in Structural BiologyThe proces

2、s and working principle of cryo - electron microscopy The advantages and limitations of cryo-Electron Microscope Application of cryo-Electron Microscope in Modern BiologyWhat is frozen electron microscope?History: 90 years of the 20th century with the frozen transmission device, field emission elect

3、ron gun and CDD imaging device appears, frozen electron microscopy single particle technology.（20世纪90年代随着冷冻传输装置、场发射电子枪以及CDD成像装置出现，冷冻电镜单颗粒技术）In the 1970s, the structure of the virus molecule was studied by using freeze electron microscopy, and the concept, method and concept of the method of freezing

4、 electron microscope were put forward for the first time（20世纪70年代通过利用冷冻电镜研究病毒分子的结构，从而首次提出冷冻电镜技术的原理，方法以及流程的概念)the early 20th century to the development of ergodic virus single particle three-dimensional reconstruction technology.（20世纪初期发展至二十面体病毒的单颗粒三维重构技术）In 2010, the three-dimensional reconstruction

5、 technique of cryo-electron microscopy was used to determine the structure of protein TRPV1, which marked that the cryostat was involved in the era of atomic resolution（2014年利用冷冻电镜三维重构技术确定蛋白质TRPV1结构，标志着冷冻电镜跨入“原子分辨率”时代）The process of cryo-electron microscopy: principleprinciple: The sample is fixed b

6、y freezing in liquid nitrogen, The sample is fixed by freezing in liquid nitrogen, making the biology large H2O molecules in the form of making the biology large H2O molecules in the form of glass in the form of existence, to maintain low glass in the form of existence, to maintain low temperature,T

7、he sample is placed in a microscope, highltemperature,The sample is placed in a microscope, highly y coherentelectrons as a light source from the above coherentelectrons as a light source from the above exposure, through the sample and the nearby ice, by exposure, through the sample and the nearby i

8、ce, by scattering, the use of detectors and lens system to record scattering, the use of detectors and lens system to record the scattered signal imaging, and then signal processing, the scattered signal imaging, and then signal processing, and finally use Three-dimensional reconstruction of the and

9、 finally use Three-dimensional reconstruction of the three-dimensional structure of the sample.three-dimensional structure of the sample.How the cryo-electron microscope works: Sample demand is lowSample demand is low closer to the physiological state closer to the physiological state Applicable to

10、a wide range of research subjects Applicable to a wide range of research subjects You can study heterogeneous samples You can study heterogeneous samples No special treatment is required for the sample No special treatment is required for the sample Dynamic snapshots of different conformations or in

11、termediaries can be Dynamic snapshots of different conformations or intermediaries can be obtainedobtained Advantages:. Expensive equipmentExpensive equipment preparation of samples difficult preparation of samples difficult samples need to be frozen, not at room temperature samples need to be froze

12、n, not at room temperature The sample may be damaged by excessive electron beam The sample may be damaged by excessive electron beamcryo-electron microscopy limitations: 1. Study the structure of molecules and their polymers that are not suitable 1. Study the structure of molecules and their polymer

13、s that are not suitable for the application of X-ray crystallography and nuclear magnetic resonance for the application of X-ray crystallography and nuclear magnetic resonance spectroscopyspectroscopySuch as: Alzhiemer disease structure of amyloid fiber polymerSuch as: Alzhiemer disease structure of

14、 amyloid fiber polymer( (研究那些不适合于应用研究那些不适合于应用X-X-射线晶体学和核磁共振波谱学的分子及其聚合物的结构射线晶体学和核磁共振波谱学的分子及其聚合物的结构）如：如：AlzhiemerAlzhiemer疾病的淀粉状蛋白纤维聚合物的结构疾病的淀粉状蛋白纤维聚合物的结构2. Study the structure of biological macromolecules in different functional 2. Study the structure of biological macromolecules in different functio

15、nal statesstatesSuch as: ion channel switchSuch as: ion channel switch（研究生物大分子处于不同功能状态时的结构研究生物大分子处于不同功能状态时的结构) )如：离子通道开关如：离子通道开关3. Provide initial molecular displacement model and initial phase for X-ray 3. Provide initial molecular displacement model and initial phase for X-ray crystallographic ana

16、lysiscrystallographic analysisSuch as: ribosome three-dimensional structureSuch as: ribosome three-dimensional structureApplication of Frozen Electron Microscope in Modern Biology:3. Provide initial molecular displacement model and initial 3. Provide initial molecular displacement model and initial phase for X-ray crystallographic analysisphase for X-ray crystallographic analysisSuch as: ribosome three-dimensional structureSuch as: ribosome three-dimensional structure( (为为X-X-射线晶体学结构解析提供初始分子置换模型及初始相位射线晶体学结构解析提供初始分子置换模型及初始相位）如：核糖体的三维结构4. Study the structure of biological macromolec