ArticleScientific reports2017
A Global Approach for Quantitative Super Resolution and Electron Microscopy on Cryo and Epoxy Sections Using Self-labeling Protein Tags.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 45 citations in OpenAlex.
- Islet ultrastructure: past achievements and future directions.Diabetologia · 2026Review
- Structure, interaction and nervous connectivity of beta cell primary cilia.Nature communications · 2024Article
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- Application of immuno- and affinity labeling with fluorescent dyes to in-resin CLEM of Epon-embedded cells.Heliyon · 2023Article
- Synchronized proinsulin trafficking reveals delayed Golgi export accompanies β-cell secretory dysfunction in rodent models of hyperglycemia.Scientific reports · 2023Article
- Article
- Nutrient Regulation of Pancreatic Islet β-Cell Secretory Capacity and Insulin Production.Biomolecules · 2022Review
- Recent Developments in Correlative Super-Resolution Fluorescence Microscopy and Electron Microscopy.Molecules and cells · 2022Review
- ER Redox Homeostasis Regulates Proinsulin Trafficking and Insulin Granule Formation in the Pancreatic Islet β-Cell.Function (Oxford, England) · 2022Article
- Reagents and models for detecting endogenous GLP1R and GIPR.EBioMedicine · 2021Review
- Article
- Sequential in vivo labeling of insulin secretory granule pools inProceedings of the National Academy of Sciences of the United States of America · 2021Article
- Molecular Evaluation of Endoplasmic Reticulum Homeostasis Meets Humoral Immunity.Trends in cell biology · 2021Review
- Easy ultrastructural insight into the internal morphology of biological specimens by Atomic Force Microscopy.Scientific reports · 2021Article
- State-of-the-art microscopy to understand islets of Langerhans: what to expect next?Immunology and cell biology · 2021Review
- Correlation of organelle dynamics between light microscopic live imaging and electron microscopic 3D architecture using FIB-SEM.Microscopy (Oxford, England) · 2021Review
- 3D FIB-SEM reconstruction of microtubule-organelle interaction in whole primary mouse β cells.The Journal of cell biology · 2021Article
- The type 2 diabetes gene product STARD10 is a phosphoinositide-binding protein that controls insulin secretory granule biogenesis.Molecular metabolism · 2020Article
- The making of insulin in health and disease.Diabetologia · 2020Review
- Chromogranin B regulates early-stage insulin granule trafficking from the Golgi in pancreatic islet β-cells.Journal of cell science · 2019Article
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Authors and funding
10 authors at 5 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Correlative light and electron microscopy (CLEM) is a powerful approach to investigate the molecular ultrastructure of labeled cell compartments. However, quantitative CLEM studies are rare, mainly due to small sample sizes and the sensitivity of fluorescent proteins to strong fixatives and contrasting reagents for EM. Here, we show that fusion of a self-labeling protein to insulin allows for the quantification of age-distinct insulin granule pools in pancreatic beta cells by a combination of super resolution and transmission electron microscopy on Tokuyasu cryosections. In contrast to fluorescent proteins like GFP organic dyes covalently bound to self-labeling proteins retain their fluorescence also in epoxy resin following high pressure freezing and freeze substitution, or remarkably even after strong chemical fixation. This enables for the assessment of age-defined granule morphology and degradation. Finally, we demonstrate that this CLEM protocol is highly versatile, being suitable for single and dual fluorescent labeling and detection of different proteins with optimal ultrastructure preservation and contrast.
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