Structural biology inside multicellular specimens using electron cryotomography

Ido Caspy; Zhexin Wang; Tanmay A.M. Bharat

doi:10.1017/S0033583525000010

Structural biology inside multicellular specimens using electron cryotomography

Published online by Cambridge University Press: 13 January 2025

and

Ido Caspy: Affiliation:
Structural Studies Division, MRC Laboratory of Molecular Biology, Cambridge, UK
Zhexin Wang: Affiliation:
Structural Studies Division, MRC Laboratory of Molecular Biology, Cambridge, UK
Tanmay A.M. Bharat*: Affiliation:
Structural Studies Division, MRC Laboratory of Molecular Biology, Cambridge, UK
*: Corresponding author: Tanmay A.M. Bharat; Email: tbharat@mrc-lmb.cam.ac.uk

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Abstract

The electron cryomicroscopy (cryo-EM) resolution revolution has shifted structural biology into a new era, enabling the routine structure determination of macromolecular complexes at an unprecedented rate. Building on this, electron cryotomography (cryo-ET) offers the potential to visualise the native three-dimensional organisation of biological specimens, from cells to tissues and even entire organisms. Despite this huge potential, the study of tissue-like multicellular specimens via cryo-ET still presents numerous challenges, wherein many steps in the workflow are being developed or in urgent need of improvement. In this review, we outline the latest techniques currently utilised for in situ imaging of multicellular specimens, while clearly enumerating their associated limitations. We consider every step in typical workflows employed by various laboratories, including sample preparation, data collection and image analysis, to highlight recent progress and showcase prominent success stories. By considering the entire structural biology workflow for multicellular specimens, we identify which future exciting developments in hardware and software could enable comprehensive in situ structural biology investigations, bringing forth a new age of discovery in molecular structural and cell biology.

Keywords

cryo-EM cryo-ET FIB-SEM focused ion beam milling in situ imaging structural biology subtomogram averaging tissue ultrastructure vitrification volume imaging

Type: Review
Information: Quarterly Reviews of Biophysics , Volume 58 , 2025 , e6

DOI: https://doi.org/10.1017/S0033583525000010 [Opens in a new window]
Copyright: © MRC Laboratory of Molecular Biology, 2025. Published by Cambridge University Press

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Structural biology inside multicellular specimens using electron cryotomography

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