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

Paul W. Tillberg, Fei Chen, Kiryl D. Piatkevich, Yongxin Zhao, C. C. Yu, Brian P. English, Linyi Gao, Anthony Martorell, Ho Jun Suk, Fumiaki Yoshida, Ellen M. Degennaro, Douglas H. Roossien, Guanyu Gong, Uthpala Seneviratne, Steven R. Tannenbaum, Robert Desimone, Dawen Cai, Edward S. Boyden

Research output: Contribution to journalArticlepeer-review

203 Citations (Scopus)

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 (∼1/470 nm) imaging of cells and mammalian tissues on conventional microscopes.

Original languageEnglish
Pages (from-to)987-992
Number of pages6
JournalNature Biotechnology
Volume34
Issue number9
DOIs
Publication statusPublished - Sep 1 2016

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

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