Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies
Author(s)English, Brian P; Gao, Linyi; Suk, Ho-Jun; Yoshida, Fumiaki; DeGennaro, Ellen M; Roossien, Douglas H; Cai, Dawen; Tillberg, Paul W.; Chen, Fei; Piatkevich, Kiryl; Zhao, Yongxin; Yu, Chih-Chieh; Martorell, Anthony; Gong, Guanyu; Seneviratne, Uthpala Indrajith; Tannenbaum, Steven R; Desimone, Robert; Boyden, Edward; ... Show more Show less
MetadataShow full item record
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.
DepartmentMassachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences; Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science; Massachusetts Institute of Technology. Media Laboratory
Nature Publishing Group
Tillberg, Paul W; Chen, Fei; Piatkevich, Kiryl D; Zhao, Yongxin; Yu, Chih-Chieh (Jay); English, Brian P; Gao, Linyi et al. “Protein-Retention Expansion Microscopy of Cells and Tissues Labeled Using Standard Fluorescent Proteins and Antibodies.” Nature Biotechnology 34, no. 9 (July 2016): 987–992. © 2016 Macmillan Publishers Limited, part of Springer Nature
Author's final manuscript