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Systematic Comparison of Commercial Uranyl‐Alternative Stains for Negative‐ and Positive‐Staining Transmission Electron Microscopy of Organic Specimens
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This study compared commercial alternatives to uranyl acetate, a toxic and radioactive staining agent used in electron microscopy. While not directly about microplastics, electron microscopy is one of the key tools researchers use to visualize and identify nanoplastics in biological tissues and environmental samples. Safer staining alternatives could make nanoplastic research more accessible and reduce hazardous waste in laboratories studying plastic contamination.
Negative- and positive-staining transmission electron microscopy (ns/psTEM) is a cornerstone of research and diagnostics, enabling nanometer-resolution analysis of organic specimens from nanoparticles to cells without requiring costly cryo-equipment. For nearly 70 years, uranyl salts like uranyl acetate (UA) have been the gold-standard ns/psTEM-stains. However, mounting safety concerns due to their high toxicity and radioactivity have led to stricter regulations and expensive licensing requirements. Consequently, there is an urgent global demand for safer, more sustainable stains that deliver uranyl-comparable, high-quality ns/psTEM. Here, the commercially available stain-alternatives UranyLess, UAR, UA-Zero, PTA, STAIN 77, Nano-W, NanoVan, and lead citrate are systematically assessed against UA. The stains are evaluated regarding their contrast, resolution, stain-distribution, and ease-of-use in ns/psTEM across a diverse sample set, including polymethylmethacrylate-nanoplastics, phosphatidylcholine-liposomes, Influenza-A viruses, globular ferritin, fibrillar pyruvate kinase amyloids, and human lung-carcinoma cell-sections. It is shown that for this variety of samples, a ready-to-use uranyl-alternative is commercially available with comparable or even superior ns/psTEM-performance to UA using an efficient staining-protocol. Furthermore, the GUIDE4U tool is developed for the fast identification of the appropriate uranyl-replacements for each sample of interest, saving ns/psTEM-users time and costs while ensuring excellent staining results for ultrastructural analysis, thereby further catalyzing the use of safer stains.
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This review rounds up the various lab techniques scientists use to find and identify tiny plastic particles (microplastics and even smaller nanoplastics) inside human and animal tissue samples, from powerful microscopes to chemical fingerprinting tools. This matters because before we can understand whether plastic particles in our bodies are actually harmful, researchers first need reliable ways to detect and measure them—and this paper helps clarify which tools work best for which situations, laying groundwork for better human health research on plastic exposure.
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Researchers reviewed labelling techniques used to track micro- and nanoplastics in environmental studies, categorizing them into fluorescent, metal, stable isotope, and radioisotope methods. The study found that fluorescent labelling works well for tracking microplastics while metal labelling is more sensitive for nanoplastics research, though a major challenge remains in developing techniques that do not alter the inherent properties of the plastic particles being studied.
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