ArticleFrontiers in cell and developmental biology2022
A versatile enhanced freeze-substitution protocol for volume electron microscopy.
Article in Frontiers in cell and developmental biology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed.
- InsP3R signaling and actomyosin-dependent mitochondrial dynamics play essential roles in mitochondrial stress-induced longevity.Nature communications · 2026Article
- Optimisation of freeze substitution protocols for the examination of malaria parasite structure by volumetric electron microscopy.Journal of microscopy · 2026Article
- InsP3R signaling mediates mitochondrial stress-induced longevity through actomyosin-dependent mitochondrial dynamics.bioRxiv : the preprint server for biology · 2025Article
- Distinct classes of 21- and 24-nt phasiRNAs suggests diverse mechanisms of biogenesis and function in rice anther development.The plant genome · 2025Article
- Article
- Spatial patterns of hepatocyte glucose flux revealed by stable isotope tracing and multi-scale microscopy.Nature communications · 2025Article
- Comparative RNA profiling identifies stage-specific phasiRNAs and coexpressed Argonaute genes in Bambusoideae and Pooideae species.The Plant cell · 2024Article
- ER-phagy drives age-onset remodeling of endoplasmic reticulum structure-function and lifespan.bioRxiv : the preprint server for biology · 2024Article
- Modes of action and potential as a peptide-based biofungicide of a plant defensin MtDef4.Molecular plant pathology · 2024Article
- Rehydration of Freeze Substituted Brain Tissue for Pre-embedding Immunoelectron Microscopy.Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada · 2023Article
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7 authors.
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Abstract
Volume electron microscopy, a powerful approach to generate large three-dimensional cell and tissue volumes at electron microscopy resolutions, is rapidly becoming a routine tool for understanding fundamental and applied biological questions. One of the enabling factors for its adoption has been the development of conventional fixation protocols with improved heavy metal staining. However, freeze-substitution with organic solvent-based fixation and staining has not realized the same level of benefit. Here, we report a straightforward approach including osmium tetroxide, acetone and up to 3% water substitution fluid (compatible with traditional or fast freeze-substitution protocols), warm-up and transition from organic solvent to aqueous 2% osmium tetroxide. Once fully hydrated, samples were processed in aqueous based potassium ferrocyanide, thiocarbohydrazide, osmium tetroxide, uranyl acetate and lead acetate before resin infiltration and polymerization. We observed a consistent and substantial increase in heavy metal staining across diverse and difficult-to-fix test organisms and tissue types, including plant tissues (
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