Cryogenic superresolution correlative light and electron microscopy of vitreous sections

Buyun Tian; Maoge Zhou; Fengping Feng; Xiaojun Xu; Pei Wang; Huiqin Luan; Wei Ji; Yanhong Xue; Tao Xu

doi:10.52601/bpr.2022.220005

Volume 8 Issue 4

Aug. 2022

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Buyun Tian, Maoge Zhou, Fengping Feng, Xiaojun Xu, Pei Wang, Huiqin Luan, Wei Ji, Yanhong Xue, Tao Xu. Cryogenic superresolution correlative light and electron microscopy of vitreous sections[J]. Biophysics Reports, 2022, 8(4): 193-204. doi: 10.52601/bpr.2022.220005

Citation:

Buyun Tian, Maoge Zhou, Fengping Feng, Xiaojun Xu, Pei Wang, Huiqin Luan, Wei Ji, Yanhong Xue, Tao Xu. Cryogenic superresolution correlative light and electron microscopy of vitreous sections[J]. Biophysics Reports, 2022, 8(4): 193-204. doi: 10.52601/bpr.2022.220005

Citation:

Buyun Tian, Maoge Zhou, Fengping Feng, Xiaojun Xu, Pei Wang, Huiqin Luan, Wei Ji, Yanhong Xue, Tao Xu. Cryogenic superresolution correlative light and electron microscopy of vitreous sections[J]. Biophysics Reports, 2022, 8(4): 193-204. doi: 10.52601/bpr.2022.220005

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Cryogenic superresolution correlative light and electron microscopy of vitreous sections

doi: 10.52601/bpr.2022.220005

Buyun Tian^{1, 2},
Maoge Zhou¹,
Fengping Feng¹,
Xiaojun Xu¹,
Pei Wang^{1, 2},
Huiqin Luan³,
Wei Ji^{1, 2
,
,},
Yanhong Xue^{1
,
,},
Tao Xu^{1, 2
,
,}

1.
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
2.
University of Chinese Academy of Sciences, Beijing 100049, China
3.
National Research Center for Rehabilitation Technical Aids, Beijing 100176, China

More Information

Corresponding author: jiwei@ibp.ac.cn (W. Ji); xueyanhong@ibp.ac.cn (Y. Xue); xutao@ibp.ac.cn (T. Xu)
Received Date: 15 February 2022
Accepted Date: 10 June 2022

Available Online: 08 November 2022

Publish Date: 31 August 2022

Abstract

Abstract

Fluorescence microscopy and electron microscopy complement each other as the former provides labelling and localisation of specific molecules and target structures while the latter possesses excellent revolving power of fine structure in context. These two techniques can combine as correlative light and electron microscopy (CLEM) to reveal the organisation of materials within the cell. Frozen hydrated sections allow microscopic observations of cellular components in situ in a near-native state and are compatible with superresolution fluorescence microscopy and electron tomography if sufficient hardware and software support is available and a well-designed protocol is followed. The development of superresolution fluorescence microscopy greatly increases the precision of fluorescence annotation of electron tomograms. Here, we provide detailed instructions on how to perform cryogenic superresolution CLEM on vitreous sections. From fluorescence-labelled cells to high pressure freezing, cryo-ultramicrotomy, cryogenic single-molecule localisation microscopy, cryogenic electron tomography and image registration, electron tomograms with features of interest highlighted by superresolution fluorescence signals are expected to be obtained.
- Cryogenic,
- Superresolution,
- Fluorescence microscopy,
- Electron microscopy,
- Correlative light and electron microscopy (CLEM),
- Electron tomography

Buyun Tian, Maoge Zhou, Fengping Feng, Xiaojun Xu, Pei Wang, Huiqin Luan, Wei Ji, Yanhong Xue and Tao Xu declare that they have no conflict of interest.
This article does not contain any studies with human or animal subjects performed by any of the authors.

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