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

• Main Authors: English, Brian P (Author), Gao, Linyi (Contributor), Suk, Ho-Jun (Contributor), Yoshida, Fumiaki (Author), DeGennaro, Ellen M (Author), Roossien, Douglas H (Author), Cai, Dawen (Author), Tillberg, Paul W. (Contributor), Chen, Fei (Contributor), Piatkevich, Kiryl (Contributor), Zhao, Yongxin (Contributor), Yu, Chih-Chieh (Contributor), Martorell, Anthony (Contributor), Gong, Guanyu (Contributor), Seneviratne, Uthpala Indrajith (Contributor), Tannenbaum, Steven R (Contributor), Desimone, Robert (Contributor), Boyden, Edward (Contributor)
• Other Authors: Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences (Contributor), Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science (Contributor), Massachusetts Institute of Technology. Media Laboratory (Contributor)
• Format: Article
• Language: English
• Published: Nature Publishing Group, 2017-04-28T20:30:04Z.
• Subjects: Article
• Online Access: Get fulltext

 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.