Ultra-stable super-resolution fluorescence cryo-microscopy for correlative light and electron cryo-microscopy |
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| Authors: | Xiaojun Xu Yanhong Xue Buyun Tian Fengping Feng Lusheng Gu Weixing Li Wei Ji " target="_blank">Tao Xu |
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| Institution: | 1.College of Life Science and Technology,Huazhong University of Science and Technology,Wuhan,China;2.National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics,Chinese Academy of Sciences,Beijing,China;3.Center for Biological Instrument Development, Core Facility for Protein Research, Institute of Biophysics,Chinese Academy of Sciences,Beijing,China;4.College of Life Science,University of Chinese Academy of Sciences,Beijing,China |
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| Abstract: | 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 super-resolution cryo-FM that exhibits excellent thermal and mechanical stability. The temperature fluctuations in 10 h are less than 0.06 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). |
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