UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy

Russell, MRG; Lerner, TR; Burden, JJ; Nkwe, DO; Pelchen-Matthews, A; Domart, M-C; Durgan, J; ... Collinson, LM; + view all (2017) 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy. Journal of Cell Science , 130 (1) pp. 278-291. 10.1242/jcs.188433. Green open access

[img]
Preview
Text
Marsh_jcs.188433.full.pdf - Published version

Download (11MB) | Preview

Abstract

The processes of life take place in multiple dimensions, but imaging these processes in even three dimensions is challenging. Here, we describe a workflow for 3D correlative light and electron microscopy (CLEM) of cell monolayers using fluorescence microscopy to identify and follow biological events, combined with serial blockface scanning electron microscopy to analyse the underlying ultrastructure. The workflow encompasses all steps from cell culture to sample processing, imaging strategy, and 3D image processing and analysis. We demonstrate successful application of the workflow to three studies, each aiming to better understand complex and dynamic biological processes, including bacterial and viral infections of cultured cells and formation of entotic cell-in-cell structures commonly observed in tumours. Our workflow revealed new insight into the replicative niche of Mycobacterium tuberculosis in primary human lymphatic endothelial cells, HIV-1 in human monocytederived macrophages, and the composition of the entotic vacuole. The broad application of this 3D CLEM technique will make it a useful addition to the correlative imaging toolbox for biomedical research.

Type: Article
Title: 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy
Open access status: An open access version is available from UCL Discovery
DOI: 10.1242/jcs.188433
Publisher version: http://dx.doi.org/10.1242/jcs.188433
Language: English
Additional information: © 2017. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
Keywords: Life Sciences & Biomedicine, Cell Biology, 3D correlative light and electron microscopy, Serial blockface SEM, Mycobacterium tuberculosis, Autophagy, HIV-1, Entosis, HIV-1 Assembly Compartments, Plasma-membrane, Array Tomography, Image Data, Mammalian-cells, In-vivo, Macrophages, Visualization, Ultrastructure, Tuberculosis
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Lab for Molecular Cell Bio MRC-UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > Institute for Global Health
URI: https://discovery.ucl.ac.uk/id/eprint/1509339
Downloads since deposit
58Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item