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1、冷冻电镜冷冻电镜.ppt.ppt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TheHistoryofApplicationofcryo-ElectronMicroscopeinStructuralBiologyTheprocessandworkingprincipleofcry
2、o-electronmicroscopyTheadvantagesandlimitationsofcryo-ElectronMicroscopeApplicationofcryo-ElectronMicroscopeinModernBiologyWhat is frozen electron microscope?cryo-cryo-electron microscopy:electron microscopy:the use of frozen fixed the use of frozen fixed technology,low temperature technology,low te
3、mperature using a transmission using a transmission electron microscope to electron microscope to observe the sample micro-observe the sample micro-technology,resulting in the technology,resulting in the structure of biological structure of biological macromoleculesmacromoleculesHistory:90 years of
4、the 20th century with the frozen transmission device,field emission electron 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 mi
5、croscopy,and the concept,method and concept of the method of freezing 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
6、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 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）Theprocessofcry
7、o-electronmicroscopy:principleprinciple:The The sample sample is is fixed fixed by by freezing freezing in in liquid liquid nitrogen,nitrogen,making making the the biology biology large large H2O H2O molecules molecules in in the the form form of of glass glass in in the the form form of of existenc
8、e,existence,to to maintain maintain low low temperature,The temperature,The sample sample is is placed placed in in a a microscope,microscope,highlhighly y coherentelectrons coherentelectrons as as a a light light source source from from the the above above exposure,exposure,through through the the
9、sample sample and and the the nearby nearby ice,ice,by by scattering,scattering,the the use use of of detectors detectors and and lens lens system system to to record record the the scattered scattered signal signal imaging,imaging,and and then then signal signal processing,processing,and and finall
10、y finally use use Three-dimensional Three-dimensional reconstruction reconstruction of of the the three-dimensional structure of the sample.three-dimensional structure of the sample.Howthecryo-electronmicroscopeworks:Sample demand is lowSample demand is low closer to the physiological state closer t
11、o the physiological state Applicable to 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 Dy
12、namic snapshots snapshots of of different different conformations conformations or or intermediaries intermediaries can can be be obtainedobtainedAdvantages:.Expensive equipmentExpensive equipment preparation of samples difficult preparation of samples difficult samples samples need need to to be be
13、 frozen,frozen,not not at at room room temperaturetemperature The The sample sample may may be be damaged damaged by by excessive excessive electron electron beambeamcryo-electronmicroscopylimitations:1.1.Study Study the the structure structure of of molecules molecules and and their their polymers
14、polymers that that are are not not suitable suitable for for the the application application of of X-ray X-ray crystallography crystallography and and nuclear nuclear magnetic magnetic resonance resonance spectroscopyspectroscopySuch as:Alzhiemer disease structure of amyloid fiber polymerSuch as:Alz
15、hiemer disease structure of 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2.2.Study Study the the structure structure of of biological biological macromolecules macromolecules in in different diffe
16、rent functional functional 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3.3.Provide Provide initial initial molecular molecular displacement displacement model model and and initial initial phase phase for for X-ray X-ray cry
17、stallographic analysiscrystallographic analysisSuch as:ribosome three-dimensional structureSuch as:ribosome three-dimensional structureApplicationofFrozenElectronMicroscopeinModernBiology:3.3.Provide Provide initial initial molecular molecular displacement displacement model model and and initial in
18、itial 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 macromolecular complexes.Such as:virus-receptor complex structure（研究生物大分子复合物的结构）如：病毒-受体复合物的结构5.Study organelles and even the structure of living cells.Such as:biological molecules in the movement of the structure and structure changes.（研究细胞器甚至是活细胞的结构)如:生物分子在运动过程中的结构和结构的变化Thanks for watching结束结束