1. Multi-color electron microscopy by element-guided identification of cells, organelles and molecules.
- Author
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Scotuzzi M, Kuipers J, Wensveen DI, de Boer P, Hagen KC, Hoogenboom JP, and Giepmans BN
- Subjects
- Animals, Color, DNA ultrastructure, Humans, Spectrometry, X-Ray Emission, Cells ultrastructure, Elements, Microscopy, Electron methods, Organelles ultrastructure
- Abstract
Cellular complexity is unraveled at nanometer resolution using electron microscopy (EM), but interpretation of macromolecular functionality is hampered by the difficulty in interpreting grey-scale images and the unidentified molecular content. We perform large-scale EM on mammalian tissue complemented with energy-dispersive X-ray analysis (EDX) to allow EM-data analysis based on elemental composition. Endogenous elements, labels (gold and cadmium-based nanoparticles) as well as stains are analyzed at ultrastructural resolution. This provides a wide palette of colors to paint the traditional grey-scale EM images for composition-based interpretation. Our proof-of-principle application of EM-EDX reveals that endocrine and exocrine vesicles exist in single cells in Islets of Langerhans. This highlights how elemental mapping reveals unbiased biomedical relevant information. Broad application of EM-EDX will further allow experimental analysis on large-scale tissue using endogenous elements, multiple stains, and multiple markers and thus brings nanometer-scale 'color-EM' as a promising tool to unravel molecular (de)regulation in biomedicine.
- Published
- 2017
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