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Zenodo
2026
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| Online toegang: | https://doi.org/10.1002/adhm.202404870 |
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| _version_ | 1866901598213505024 |
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| author | Kissling, Dr. sc. Vera Maria Eitner, Stephanie Bottone, Davide Cereghetti, Gea Wick, Peter |
| author_facet | Kissling, Dr. sc. Vera Maria Eitner, Stephanie Bottone, Davide Cereghetti, Gea Wick, Peter |
| contents | <p><span dir="ltr">Negative- and positive-staining transmission electron microscopy </span><span dir="ltr">(ns/psTEM) is a cornerstone of research and diagnostics, enabling </span><span dir="ltr">nanometer-resolution analysis of organic specimens from nanoparticles to </span><span dir="ltr">cells without requiring costly cryo-equipment. For nearly 70 years, uranyl salts </span><span dir="ltr">like uranyl acetate (UA) have been the gold-standard ns/psTEM-stains.</span><br><span dir="ltr">However, mounting safety concerns due to their high toxicity and radioactivity </span><span dir="ltr">have led to stricter regulations and expensive licensing requirements. </span><span dir="ltr">Consequently, there is an urgent global demand for safer, more sustainable </span><span dir="ltr">stains that deliver uranyl-comparable, high-quality ns/psTEM. Here, the </span><span dir="ltr">commercially available stain-alternatives UranyLess, UAR, UA-Zero, PTA, </span><span dir="ltr">STAIN 77, Nano-W, NanoVan, and lead citrate are systematically assessed </span><span dir="ltr">against UA. The stains are evaluated regarding their contrast, resolution, </span><span dir="ltr">stain-distribution, and ease-of-use in ns/psTEM across a diverse sample set, </span><span dir="ltr">including polymethylmethacrylate-nanoplastics, </span><span dir="ltr">phosphatidylcholine-liposomes, Influenza-A viruses, globular ferritin, fibrillar </span><span dir="ltr">pyruvate kinase amyloids, and human lung-carcinoma cell-sections. It is </span><span dir="ltr">shown that for this variety of samples, a ready-to-use uranyl-alternative is </span><span dir="ltr">commercially available with comparable or even superior </span><span dir="ltr">ns/psTEM-performance to UA using an efficient staining-protocol.</span><br><span dir="ltr">Furthermore, the</span> <span dir="ltr">GUIDE4U</span> <span dir="ltr">tool is developed for the fast identification of the </span><span dir="ltr">appropriate uranyl-replacements for each sample of interest, saving </span><span dir="ltr">ns/psTEM-users time and costs while ensuring excellent staining results for </span><span dir="ltr">ultrastructural analysis, thereby further catalyzing the use of safer stains.</span></p> <p> </p> <p><span dir="ltr">Acknowledgements:</span></p> <p><span dir="ltr"><span dir="ltr">Work in the </span><span dir="ltr">Nanomaterial in Health Laboratory profited from funding by Empa and </span><span dir="ltr">the MetrINo project (23.00360, 22HLT04, received funding from the Eu</span><span dir="ltr">ropean Partnership on Metrology, co-financed by the European Union’s </span><span dir="ltr">Horizon Europe Research and Innovation Programme and SERI (REF-</span><span dir="ltr">1131-52104)). Dr. Gea Cereghetti was supported by an EMBO Fellowship </span><span dir="ltr">(EMBO ALTF 349–2023), and a UKRI Engineering and Physical Sciences </span><span dir="ltr">Research Council Fellowship (grant EP/Z000033/1).</span></span></p> |
| format | Recurso digital |
| id | zenodo_https___doi_org_10_1002_adhm_202404870 |
| institution | Zenodo |
| language | |
| publishDate | 2026 |
| publisher | Zenodo |
| record_format | zenodo |
| spellingShingle | Systematic Comparison of Commercial Uranyl-Alternative Stains for Negative- and Positive-Staining Transmission Electron Microscopy of Organic Specimens Kissling, Dr. sc. Vera Maria Eitner, Stephanie Bottone, Davide Cereghetti, Gea Wick, Peter <p><span dir="ltr">Negative- and positive-staining transmission electron microscopy </span><span dir="ltr">(ns/psTEM) is a cornerstone of research and diagnostics, enabling </span><span dir="ltr">nanometer-resolution analysis of organic specimens from nanoparticles to </span><span dir="ltr">cells without requiring costly cryo-equipment. For nearly 70 years, uranyl salts </span><span dir="ltr">like uranyl acetate (UA) have been the gold-standard ns/psTEM-stains.</span><br><span dir="ltr">However, mounting safety concerns due to their high toxicity and radioactivity </span><span dir="ltr">have led to stricter regulations and expensive licensing requirements. </span><span dir="ltr">Consequently, there is an urgent global demand for safer, more sustainable </span><span dir="ltr">stains that deliver uranyl-comparable, high-quality ns/psTEM. Here, the </span><span dir="ltr">commercially available stain-alternatives UranyLess, UAR, UA-Zero, PTA, </span><span dir="ltr">STAIN 77, Nano-W, NanoVan, and lead citrate are systematically assessed </span><span dir="ltr">against UA. The stains are evaluated regarding their contrast, resolution, </span><span dir="ltr">stain-distribution, and ease-of-use in ns/psTEM across a diverse sample set, </span><span dir="ltr">including polymethylmethacrylate-nanoplastics, </span><span dir="ltr">phosphatidylcholine-liposomes, Influenza-A viruses, globular ferritin, fibrillar </span><span dir="ltr">pyruvate kinase amyloids, and human lung-carcinoma cell-sections. It is </span><span dir="ltr">shown that for this variety of samples, a ready-to-use uranyl-alternative is </span><span dir="ltr">commercially available with comparable or even superior </span><span dir="ltr">ns/psTEM-performance to UA using an efficient staining-protocol.</span><br><span dir="ltr">Furthermore, the</span> <span dir="ltr">GUIDE4U</span> <span dir="ltr">tool is developed for the fast identification of the </span><span dir="ltr">appropriate uranyl-replacements for each sample of interest, saving </span><span dir="ltr">ns/psTEM-users time and costs while ensuring excellent staining results for </span><span dir="ltr">ultrastructural analysis, thereby further catalyzing the use of safer stains.</span></p> <p> </p> <p><span dir="ltr">Acknowledgements:</span></p> <p><span dir="ltr"><span dir="ltr">Work in the </span><span dir="ltr">Nanomaterial in Health Laboratory profited from funding by Empa and </span><span dir="ltr">the MetrINo project (23.00360, 22HLT04, received funding from the Eu</span><span dir="ltr">ropean Partnership on Metrology, co-financed by the European Union’s </span><span dir="ltr">Horizon Europe Research and Innovation Programme and SERI (REF-</span><span dir="ltr">1131-52104)). Dr. Gea Cereghetti was supported by an EMBO Fellowship </span><span dir="ltr">(EMBO ALTF 349–2023), and a UKRI Engineering and Physical Sciences </span><span dir="ltr">Research Council Fellowship (grant EP/Z000033/1).</span></span></p> |
| title | Systematic Comparison of Commercial Uranyl-Alternative Stains for Negative- and Positive-Staining Transmission Electron Microscopy of Organic Specimens |
| url | https://doi.org/10.1002/adhm.202404870 |