title: Correlative fluorescence and scanning transmission electron microscopy of quantum dot-labeled proteins on whole cells in liquid.
abstract: Correlative fluorescence microscopy combined with scanning transmission electron microscopy (STEM) of cells fully immersed in liquid is a new methodology with many application areas. Proteins, in live cells immobilized on microchips, are labeled with fluorescent quantum dot nanoparticles. In this protocol, the epidermal growth factor receptor (EGFR) is labeled. The cells are fixed after a selected labeling time, for example, 5 min as needed to form EGFR dimers. The microchip with cells is then imaged with fluorescence microscopy. Thereafter, STEM can be accomplished in two ways. The microchip with the labeled cells and one microchip with a spacer are assembled into a special microfluidic device and imaged with dedicated high-voltage STEM. Alternatively, thin edges of cells can be studied with environmental scanning electron microscopy with a STEM detector, by placing a microchip with cells in a cooled wet environment.
authors.affiliation: Innovative Electron Microscopy, INM-Leibniz Institute for New Materials, Saarbrücken, Germany.
authors.forename: Diana B
authors.initials: DB
authors.lastname: Peckys
authors.affiliation: Biomineralization, INM-Leibniz Institute for New Materials, Saarbrücken, Germany.
authors.forename: Vera
authors.initials: V
authors.lastname: Bandmann
authors.affiliation: Innovative Electron Microscopy, INM-Leibniz Institute for New Materials, Saarbrücken, Germany.
authors.forename: Niels
authors.initials: N
authors.lastname: de Jonge
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D066246:ErbB Receptors / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry*; D017348:Microscopy, Electron, Scanning Transmission / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D045663:Quantum Dots / Q000737:chemistry*; D059010:Single-Cell Analysis; D013194:Staining and Labeling
chemical_list: D005456:Fluorescent Dyes; D066246:ErbB Receptors
keywords: ESEM; Environmental scanning electron microscopy; Epidermal growth factor receptor; Fluorescence microscopy; Quantum dot; STEM; Scanning transmission electron microscopy; Specific label; Whole cell