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

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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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Acknowledgements

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

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  1. Contributed equally to this work

Authors and Affiliations

  1. College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China

    Xiaojun Xu & Tao Xu

  2. National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China

    Xiaojun Xu, Yanhong Xue, Fengping Feng & Tao Xu

  3. Center for Biological Instrument Development, Core Facility for Protein Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China

    Lusheng Gu, Weixing Li, Wei Ji & Tao Xu

  4. College of Life Science, University of Chinese Academy of Sciences, Beijing, 100049, China

    Buyun Tian, Wei Ji & Tao Xu

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  1. Xiaojun Xu
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  2. Yanhong Xue
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  3. Buyun Tian
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  4. Fengping Feng
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  7. Wei Ji
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  8. Tao Xu
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Correspondence to Wei Ji or Tao Xu.

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Xu, X., Xue, Y., Tian, B. et al. Ultra-stable super-resolution fluorescence cryo-microscopy for correlative light and electron cryo-microscopy. Sci. China Life Sci. 61, 1312–1319 (2018). https://doi.org/10.1007/s11427-018-9380-3

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  • DOI: https://doi.org/10.1007/s11427-018-9380-3

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