FAST-EM array tomography: a workflow for multibeam volume electron microscopy

Publication date

2024-04

Authors

Kievits, Arent J
Duinkerken, B H Peter
Lane, Ryan
de Heus, Cecilia
van Beijeren Bergen En Henegouwen, Daan
Höppener, Tibbe
Wolters, Anouk H G
Liv, NalanORCID 0000-0003-2654-9117
Giepmans, Ben N G
Hoogenboom, Jacob P

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Document Type

Article

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cc_by

Abstract

Elucidating the 3D nanoscale structure of tissues and cells is essential for understanding the complexity of biological processes. Electron microscopy (EM) offers the resolution needed for reliable interpretation, but the limited throughput of electron microscopes has hindered its ability to effectively image large volumes. We report a workflow for volume EM with FAST-EM, a novel multibeam scanning transmission electron microscope that speeds up acquisition by scanning the sample in parallel with 64 electron beams. FAST-EM makes use of optical detection to separate the signals of the individual beams. The acquisition and 3D reconstruction of ultrastructural data from multiple biological samples is demonstrated. The results show that the workflow is capable of producing large reconstructed volumes with high resolution and contrast to address biological research questions within feasible acquisition time frames.

Keywords

volume electron microscopy, FAST-EM, array tomography, image processing, automatic segmentation, Journal Article

Citation

Kievits, A J, Duinkerken, B H P, Lane, R, de Heus, C, van Beijeren Bergen En Henegouwen, D, Höppener, T, Wolters, A H G, Liv, N, Giepmans, B N G & Hoogenboom, J P 2024, 'FAST-EM array tomography : a workflow for multibeam volume electron microscopy', Methods in microscopy, vol. 1, no. 1, pp. 49-64. https://doi.org/10.1515/mim-2024-0005