title: Electron Microscopy of Living Cells During in Situ Fluorescence Microscopy.
abstract: We present an approach toward dynamic nanoimaging: live fluorescence of cells encapsulated in a bionanoreactor is complemented with in situ scanning electron microscopy (SEM) on an integrated microscope. This allows us to take SEM snapshots on-demand, that is, at a specific location in time, at a desired region of interest, guided by the dynamic fluorescence imaging. We show that this approach enables direct visualization, with EM resolution, of the distribution of bioconjugated quantum dots on cellular extensions during uptake and internalization.
authors.affiliation: Department of Imaging Physics, Delft University of Technology , Lorentzweg 1, 2628 CJ Delft, The Netherlands.
authors.forename: Nalan
authors.initials: N
authors.lastname: Liv
authors.affiliation: Department of Imaging Physics, Delft University of Technology , Lorentzweg 1, 2628 CJ Delft, The Netherlands.
authors.forename: Daan S B
authors.initials: DS
authors.lastname: van Oosten Slingeland
authors.affiliation: Department of Molecular Physiology and Biophysics, Vanderbilt University , 747 Light Hall, Nashville, Tennessee 37232-0615, United States.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Baudoin
authors.affiliation: Department of Imaging Physics, Delft University of Technology , Lorentzweg 1, 2628 CJ Delft, The Netherlands.
authors.forename: Pieter
authors.initials: P
authors.lastname: Kruit
authors.affiliation: Department of Molecular Physiology and Biophysics, Vanderbilt University , 747 Light Hall, Nashville, Tennessee 37232-0615, United States.
authors.forename: David W
authors.initials: DW
authors.lastname: Piston
authors.affiliation: Department of Imaging Physics, Delft University of Technology , Lorentzweg 1, 2628 CJ Delft, The Netherlands.
authors.forename: Jacob P
authors.initials: JP
authors.lastname: Hoogenboom
mesh_terms: D000818:Animals; D019149:Bioreactors; D002460:Cell Line; D018914:Cells, Immobilized / Q000378:metabolism / Q000648:ultrastructure*; D002522:Chlorocebus aethiops; D004705:Endocytosis / Q000502:physiology; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure*; D008855:Microscopy, Electron, Scanning / Q000295:instrumentation / Q000379:methods*; D061848:Optical Imaging / Q000295:instrumentation / Q000379:methods*; D045663:Quantum Dots / Q000737:chemistry / Q000378:metabolism
keywords: epidermal growth factor; fluorescence microscopy; live cell imaging; quantum dots; scanning electron microscopy