Ultra-stable super-resolution fluorescence cryo-microscopy for correlative light and electron cryo-microscopy  

作　　者：Xiaojun Xu Yanhong Xue Buyun Tian Fengping Feng Lusheng Gu Weixing Li Wei Ji Tao Xu 

机构地区：[1]College of Life Science and Technology,Huazhong University of Science and Technology,Wuhan 430074,China [2]National Laboratoty of Biomacromolecules,CAS Center for Excellence in Biomacromolecules,Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China [3]Center for Biological Instrument Development,Core Facility for Protein Research,Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China [4]College of Life Science,University of Chinese Academy of Sciences,Beijing 100049 China

出　　处：《Science China(Life Sciences)》2018年第11期1312-1319,共8页中国科学（生命科学英文版）

基　　金：supported by the National Key R&D Program of China (2016YFA0500203, 2016YFA0502400, 2017YFA0504700, 2017YFA0505300);the National Natural Science Foundation of China (31661143041, 31127901);Joint Program between Chinese Academy of Sciences and Peking University

摘　　要：Remarkable progress in correlative light and electron cryo-microscopy(cryo-CLEM) has been made in the past decade. A crucial component for cryo-CLEM is a dedicated cryo-fluorescence microscope(cryo-FM). Here, we describe an ultra-stable superresolution cryo-FM that exhibits excellent thermal and mechanical stability. The temperature fluctuations in 10 h are less than0.06 K, and the mechanical drift over 5 h is less than 200 nm in three dimensions. We have demonstrated the super-resolution imaging capability of this system(average single molecule localization accuracy of ～13.0 nm). The results suggest that our system is particularly suitable for long-term observations, such as single molecule localization microscopy(SMLM) and cryogenic super-resolution correlative light and electron microscopy(csCLEM).Remarkable progress in correlative light and electron cryo-microscopy(cryo-CLEM) has been made in the past decade. A crucial component for cryo-CLEM is a dedicated cryo-fluorescence microscope(cryo-FM). Here, we describe an ultra-stable superresolution cryo-FM that exhibits excellent thermal and mechanical stability. The temperature fluctuations in 10 h are less than0.06 K, and the mechanical drift over 5 h is less than 200 nm in three dimensions. We have demonstrated the super-resolution imaging capability of this system(average single molecule localization accuracy of ～13.0 nm). The results suggest that our system is particularly suitable for long-term observations, such as single molecule localization microscopy(SMLM) and cryogenic super-resolution correlative light and electron microscopy(csCLEM).

关 键 词：CRYOGENIC CRYOSTAT SUPER-RESOLUTION Cryo-FM Cryo-PALM csCLEM 

分 类 号：Q[生物学]