title: Fixation-resistant photoactivatable fluorescent proteins for CLEM.
abstract: Fluorescent proteins facilitate a variety of imaging paradigms in live and fixed samples. However, they lose their fluorescence after heavy fixation, hindering applications such as correlative light and electron microscopy (CLEM). Here we report engineered variants of the photoconvertible Eos fluorescent protein that fluoresce and photoconvert normally in heavily fixed (0.5-1% OsO4), plastic resin-embedded samples, enabling correlative super-resolution fluorescence imaging and high-quality electron microscopy.
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Maria G
authors.initials: MG
authors.lastname: Paez-Segala
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Mei G
authors.initials: MG
authors.lastname: Sun
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Gleb
authors.initials: G
authors.lastname: Shtengel
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Sarada
authors.initials: S
authors.lastname: Viswanathan
authors.affiliation: 1] National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA. [2] Department of Biological Science, Florida State University, Tallahassee, Florida, USA.
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Baird
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: John J
authors.initials: JJ
authors.lastname: Macklin
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Ronak
authors.initials: R
authors.lastname: Patel
authors.affiliation: 1] National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA. [2] Department of Biological Science, Florida State University, Tallahassee, Florida, USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Allen
authors.affiliation: 1] National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA. [2] Department of Biological Science, Florida State University, Tallahassee, Florida, USA.
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: Howe
authors.affiliation: National Heart, Lung, and Blood Institute, Bethesda, Maryland, USA.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Piszczek
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Harald F
authors.initials: HF
authors.lastname: Hess
authors.affiliation: 1] National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA. [2] Department of Biological Science, Florida State University, Tallahassee, Florida, USA.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Davidson
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Yalin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Loren L
authors.initials: LL
authors.lastname: Looger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D003412:Cricetulus; D005453:Fluorescence; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods; D008969:Molecular Sequence Data; D009993:Osmium Tetroxide / Q000737:chemistry; D010777:Photochemistry / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D009993:Osmium Tetroxide