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Nat Biotechnol. 2016 Sep;34(9):987-92. doi: 10.1038/nbt.3625. Epub 2016 Jul 4.

Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies.

Author information

1
Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
2
Massachusetts Institute of Technology Media Lab, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
3
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
4
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.
5
Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
6
Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
7
Osaka University Medical School, Suita, Osaka, Japan.
8
McGovern Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
9
University of Michigan Medical School, Ann Arbor, Michigan, USA.

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.

Comment in

PMID:
27376584
PMCID:
PMC5068827
DOI:
10.1038/nbt.3625
[Indexed for MEDLINE]
Free PMC Article

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