Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies
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...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Nature Publishing Group
2017
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| Online Access: | http://hdl.handle.net/1721.1/108514 https://orcid.org/0000-0003-0254-4741 https://orcid.org/0000-0003-3776-4605 https://orcid.org/0000-0003-4188-5725 https://orcid.org/0000-0001-8713-0446 https://orcid.org/0000-0002-3579-0327 https://orcid.org/0000-0002-2206-2590 https://orcid.org/0000-0002-0899-6709 https://orcid.org/0000-0001-6774-9639 https://orcid.org/0000-0002-5938-4227 https://orcid.org/0000-0002-0419-3351 |
| Summary: | 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. |
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