title: Correlative Fluorescence and Electron Microscopy in 3D-Scanning Electron Microscope Perspective.
abstract: The ability to correlate fluorescence microscopy (FM) and electron microscopy (EM) data obtained on biological (cell and tissue) specimens is essential to bridge the resolution gap between the data obtained by these different imaging techniques. In the past such correlations were limited to either EM navigation in two dimensions to the locations previously highlighted by fluorescence markers, or subsequent high-resolution acquisition of tomographic information using a TEM. We present a novel approach whereby a sample previously investigated by FM is embedded and subjected to sequential mechanical polishing and backscatter imaging by scanning electron microscope. The resulting three dimensional EM tomogram of the sample can be directly correlated to the FM data. © 2017 by John Wiley & Sons, Inc.
authors.affiliation: Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Franks
authors.affiliation: Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Callen T
authors.initials: CT
authors.lastname: Wallace
authors.affiliation: JEOL USA Inc, Peabody, Massachusetts.
authors.forename: Masateru
authors.initials: M
authors.lastname: Shibata
authors.affiliation: JEOL Ltd, Akishima, Tokyo, Japan.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Suga
authors.affiliation: JEOL USA Inc, Peabody, Massachusetts.
authors.forename: Natasha
authors.initials: N
authors.lastname: Erdman
authors.affiliation: Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Donna B
authors.initials: DB
authors.lastname: Stolz
authors.affiliation: Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Simon C
authors.initials: SC
authors.lastname: Watkins
mesh_terms: D006367:HeLa Cells; D006801:Humans; D021621:Imaging, Three-Dimensional; D007668:Kidney / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*
keywords: EM tomography; SEM; fluorescence; low vacuum; mechanical polishing