title: Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies.
abstract: Expansion microscopy (ExM) enables imaging of preserved specimens with nanoscale precision on diffraction-limited instead of specialized super-resolution microscopes. ExM works by physically separating fluorescent probes after anchoring them to a swellable gel. The first ExM method did not result in the retention of native proteins in the gel and relied on custom-made reagents that are not widely available. Here we describe protein retention ExM (proExM), a variant of ExM in which proteins are anchored to the swellable gel, allowing the use of conventional fluorescently labeled antibodies and streptavidin, and fluorescent proteins. We validated and demonstrated the utility of proExM for multicolor super-resolution (∼70 nm) imaging of cells and mammalian tissues on conventional microscopes.
authors.affiliation: Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Massachusetts Institute of Technology Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Tillberg
authors.affiliation: Massachusetts Institute of Technology Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Fei
authors.initials: F
authors.lastname: Chen
authors.affiliation: Massachusetts Institute of Technology Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Kiryl D
authors.initials: KD
authors.lastname: Piatkevich
authors.affiliation: Massachusetts Institute of Technology Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Yongxin
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Massachusetts Institute of Technology Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Chih-Chieh Jay
authors.initials: CC
authors.lastname: Yu
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.
authors.forename: Brian P
authors.identifier: 0000-0002-4037-6294
authors.initials: BP
authors.lastname: English
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Linyi
authors.identifier: 0000-0002-3579-0327
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Anthony
authors.initials: A
authors.lastname: Martorell
authors.affiliation: Massachusetts Institute of Technology Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Ho-Jun
authors.initials: HJ
authors.lastname: Suk
authors.affiliation: Osaka University Medical School, Suita, Osaka, Japan.|McGovern Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Yoshida
authors.affiliation: Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|McGovern Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Ellen M
authors.identifier: 0000-0001-5506-6451
authors.initials: EM
authors.lastname: DeGennaro
authors.affiliation: University of Michigan Medical School, Ann Arbor, Michigan, USA.
authors.forename: Douglas H
authors.initials: DH
authors.lastname: Roossien
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Guanyu
authors.initials: G
authors.lastname: Gong
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Uthpala
authors.initials: U
authors.lastname: Seneviratne
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Tannenbaum
authors.affiliation: Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|McGovern Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Desimone
authors.affiliation: University of Michigan Medical School, Ann Arbor, Michigan, USA.
authors.forename: Dawen
authors.initials: D
authors.lastname: Cai
authors.affiliation: Massachusetts Institute of Technology Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|McGovern Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Edward S
authors.initials: ES
authors.lastname: Boyden
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000166:cytology* / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D007089:Image Enhancement / Q000379:methods*; D008164:Luminescent Proteins; D008253:Macaca mulatta; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D008164:Luminescent Proteins