ArticleScientific reports2020
Two-color in-resin CLEM of Epon-embedded cells using osmium resistant green and red fluorescent proteins.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 29 citations in OpenAlex.
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- Diverse applications of volume CLEM analysis in cell biology: brief overview, unique techniques and recent notable works.Microscopy (Oxford, England) · 2026Review
- Advances in multiscale myelin imaging: from classical histology to functional insights.Frontiers in cellular neuroscience · 2026Review
- Multicolor cathodoluminescence imaging of single lanthanide nanoparticles.Nature communications · 2025Article
- α-Synuclein-Assembled Synaptic Vesicle Pools at the Presynaptic Terminal: A Study of α-Synuclein Function Using a Novel Mouse Model.Acta histochemica et cytochemica · 2025Review
- A highly stable monomeric red fluorescent protein for advanced microscopy.Nature methods · 2025Article
- Article
- Application of immuno- and affinity labeling with fluorescent dyes to in-resin CLEM of Epon-embedded cells.Heliyon · 2023Article
- In-resin CLEM of Epon-embedded cells using proximity labeling.Scientific reports · 2022Article
- Article
- Multi-scale light microscopy/electron microscopy neuronal imaging from brain to synapse with a tissue clearing method, ScaiScience · 2022Article
- Recent technological advances in correlative light and electron microscopy for the comprehensive analysis of neural circuits.Frontiers in neuroanatomy · 2022Review
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In-resin CLEM of Epon embedded samples can greatly simplify the correlation of fluorescent images with electron micrographs. The usefulness of this technique is limited at present by the low number of fluorescent proteins that resist CLEM processing. Additionally, no study has reported the possibility of two-color in-resin CLEM of Epon embedded cells. In this study, we screened for monomeric green and red fluorescent proteins that resist CLEM processing. We identified mWasabi, CoGFP variant 0, and mCherry2; two green and one red fluorescent proteins as alternatives for in-resin CLEM. We expressed mitochondria-localized mCherry2 and histone H2B tagged with CoGFP variant 0 in cells. Green and red fluorescence was detected in 100 nm-thin sections of the Epon-embedded cells. In the same thin sections, we correlated the fluorescent signals to mitochondria and the nucleus using a scanning electron microscope. Similar results were obtained when endoplasmic reticulum-localized mCherry2 and histone H2B tagged with CoGFP variant 0 were expressed in the cells. Two-color in-resin CLEM of two cytoplasmic organelles, mitochondria and endoplasmic reticulum, was also achieved using mitochondria-localized mCherry2 and endoplasmic reticulum-localized mWasabi. In summary, we report three new fluorescent protein-alternatives suitable for in-resin CLEM of Epon-embedded samples, and achieved Epon-based two-color in-resin CLEM.
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