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Sample Processing and Benchmarking for Multibeam Optical Scanning Transmission Electron Microscopy
B H Peter Duinkerken1, Arent J Kievits2, Anouk H G Wolters1
1Department of Biomedical Sciences, University of Groningen, University Medical Centre Groningen, Antonius Deusinglaan 1, Groningen, AV 9713, The Netherlands.
Summary
Multibeam optical scanning transmission electron microscopy (OSTEM) offers an order of magnitude speed increase over conventional methods. This high-throughput technique provides comparable image quality for biological ultrastructure analysis, enabling larger-scale studies.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biotechnology
Background:
- Electron microscopy (EM) is crucial for visualizing biological ultrastructure.
- High-throughput EM is needed for analyzing larger biological samples and volumes.
- Multibeam optical scanning transmission EM (OSTEM) promises increased imaging throughput.
Purpose of the Study:
- To evaluate the compatibility of multibeam OSTEM with standard sample preparation.
- To determine the impact of machine settings on multibeam OSTEM image quality.
- To compare the speed and quality of multibeam OSTEM with conventional EM techniques.
Main Methods:
- Utilized multibeam OSTEM with multiple beamlets and optical electron separation.
- Employed standard high-contrast staining protocols for biological tissue samples.
- Investigated optimal acceleration voltage (5 kV), section thickness, and pixel dwell time.
Main Results:
- Multibeam OSTEM achieved an order of magnitude higher throughput than conventional EM.
- Generated high-quality images comparable to standard transmission EM modalities.
- Demonstrated flexibility in stain types, with optimal results using embedding methods.
Conclusions:
- Multibeam OSTEM is a viable high-throughput EM technique for ultrastructure analysis.
- Achieves comparable image quality to traditional methods while significantly increasing speed.
- Enables analysis of biological ultrastructure across larger scales and volumes.
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