Multi-color electron microscopy by element-guided identification of cells, organelles and molecules

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 complement...

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Published inScientific reports Vol. 7; no. 1; p. 45970
Main Authors Scotuzzi, Marijke, Kuipers, Jeroen, Wensveen, Dasha I., de Boer, Pascal, Hagen, Kees (C.) W., Hoogenboom, Jacob P., Giepmans, Ben N. G.
Format Journal Article
LanguageEnglish
Published London Nature Publishing Group UK 07.04.2017
Nature Publishing Group
Subjects
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14/1
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631/1647/328
631/80/642
Antibodies
Cadmium
Chemical composition
Color
Data analysis
Data processing
Diabetes
Electron microscopy
Endoplasmic reticulum
Glucagon
Humanities and Social Sciences
Identification
Insulin
Islets of Langerhans
Localization
Macromolecules
Mapping
Microscopy
multidisciplinary
Nanoparticles
Organelles
Pancreas
Quantum dots
Science
Spectrum analysis
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Summary: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.
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These authors contributed equally to this work.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep45970