title: Multi-color electron microscopy by element-guided identification of cells, organelles and molecules.
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.
authors.affiliation: Dept. Imaging Physics, Delft University of Technology, Delft, The Netherlands.
authors.forename: Marijke
authors.initials: M
authors.lastname: Scotuzzi
authors.affiliation: Dept. Cell Biology, University Medical Centre Groningen, University of Groningen, Groningen, The Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Kuipers
authors.affiliation: Dept. Imaging Physics, Delft University of Technology, Delft, The Netherlands.
authors.forename: Dasha I
authors.initials: DI
authors.lastname: Wensveen
authors.affiliation: Dept. Cell Biology, University Medical Centre Groningen, University of Groningen, Groningen, The Netherlands.
authors.forename: Pascal
authors.initials: P
authors.lastname: de Boer
authors.affiliation: Dept. Imaging Physics, Delft University of Technology, Delft, The Netherlands.
authors.forename: Kees C W
authors.initials: KC
authors.lastname: Hagen
authors.affiliation: Dept. Imaging Physics, Delft University of Technology, Delft, The Netherlands.
authors.forename: Jacob P
authors.initials: JP
authors.lastname: Hoogenboom
authors.affiliation: Dept. Cell Biology, University Medical Centre Groningen, University of Groningen, Groningen, The Netherlands.
authors.forename: Ben N G
authors.initials: BN
authors.lastname: Giepmans
mesh_terms: D000818:Animals; D002477:Cells / Q000648:ultrastructure*; D003116:Color; D004247:DNA / Q000648:ultrastructure; D004602:Elements; D006801:Humans; D008854:Microscopy, Electron / Q000379:methods*; D015388:Organelles / Q000648:ultrastructure*; D013052:Spectrometry, X-Ray Emission
chemical_list: D004602:Elements; D004247:DNA