FAST-EM array tomography:
a workflow for multibeam volume electron microscopy
A.J. Kievits (TU Delft - Applied Sciences)
B.H. Peter Duinkerken (University Medical Center Groningen)
R. Lane (TU Delft - Applied Sciences)
Cecilia de Heus ( University Medical Centre Utrecht)
Daan van Beijeren Bergen en Henegouwen (University Medical Center Groningen)
T.R. Höppener (TU Delft - Applied Sciences)
Anouk H.G. Wolters (University Medical Center Groningen)
Nalan Liv ( University Medical Centre Utrecht)
Ben N.G. Giepmans (University Medical Center Groningen)
J.P. Hoogenboom (TU Delft - Applied Sciences)
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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.
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