ArticleJournal of microscopy2018
High-performance serial block-face SEM of nonconductive biological samples enabled by focal gas injection-based charge compensation.
Article in Journal of microscopy, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 60 papers.
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Who cites it
60 citing papers in PubMed.
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- Multiscale mitochondrial cristae remodeling links Opa1 downregulation to reduced OXPHOS capacity in aged hearts.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Daily ultrastructural remodeling of clock neurons.Current biology : CB · 2025Article
- Biological volume EM with focused Ga ion beam depends on formation of radiation-resistant Ga-rich layer at block face.Research square · 2025Article
- Reduction of SEM charging artefacts in native cryogenic biological samples.Nature communications · 2025Article
- New Approaches Based on Serial-Block Face Electron Microscopy to Investigate the Peripheral Nervous System.Journal of the peripheral nervous system : JPNS · 2025Article
- Advancing yeast cell analysis: A cryomethod for serial block-face scanning electron microscopy imaging in mitochondrial morphology studies.Biology of the cell · 2025Article
- Repetitive ultramicrotome trimming and SEM imaging for characterizing printed multilayer structures.Scientific reports · 2024Article
- Protocol for volume correlative light X-ray and electron microscopy of endothelial cells in mouse tissue.STAR protocols · 2024Article
- Sample preparation and data collection for serial block face scanning electron microscopy of mammalian cell monolayers.PloS one · 2024Article
- A Comprehensive Approach to Sample Preparation for Electron Microscopy and the Assessment of Mitochondrial Morphology in Tissue and Cultured Cells.Advanced biology · 2023Article
- Piccolino is required for ribbon architecture at cochlear inner hair cell synapses and for hearing.EMBO reports · 2023Article
- Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest.Frontiers in zoology · 2023Article
- Astrocytic engagement of the corticostriatal synaptic cleft is disrupted in a mouse model of Huntington's disease.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Phosphorylation of PSD-95 at serine 73 in dCA1 is required for extinction of contextual fear.PLoS biology · 2023Article
- De novo evolution of macroscopic multicellularity.Nature · 2023Article
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Authors and funding
6 authors.
Funding
Abstract
A longstanding limitation of imaging with serial block-face scanning electron microscopy is specimen surface charging. This charging is largely due to the difficulties in making biological specimens and the resins in which they are embedded sufficiently conductive. Local accumulation of charge on the specimen surface can result in poor image quality and distortions. Even minor charging can lead to misalignments between sequential images of the block-face due to image jitter. Typically, variable-pressure SEM is used to reduce specimen charging, but this results in a significant reduction to spatial resolution, signal-to-noise ratio and overall image quality. Here we show the development and application of a simple system that effectively mitigates specimen charging by using focal gas injection of nitrogen over the sample block-face during imaging. A standard gas injection valve is paired with a precisely positioned but retractable application nozzle, which is mechanically coupled to the reciprocating action of the serial block-face ultramicrotome. This system enables the application of nitrogen gas precisely over the block-face during imaging while allowing the specimen chamber to be maintained under high vacuum to maximise achievable SEM image resolution. The action of the ultramicrotome drives the nozzle retraction, automatically moving it away from the specimen area during the cutting cycle of the knife. The device described was added to a Gatan 3View system with minimal modifications, allowing high-resolution block-face imaging of even the most charge prone of epoxy-embedded biological samples.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.