{"entity": "publication", "iuid": "d424cf3383cb4f00b90f3c15f327aeae", "timestamp": "2026-06-18T15:01:27.430Z", "links": {"self": {"href": "https://publications.scilifelab.se/publication/d424cf3383cb4f00b90f3c15f327aeae.json"}, "display": {"href": "https://publications.scilifelab.se/publication/d424cf3383cb4f00b90f3c15f327aeae"}}, "title": "Dual-Color Expansion Microscopy of Membrane Proteins Using Bioorthogonal Labeling", "authors": [{"family": "Edwards", "given": "Steven", "initials": "S", "orcid": "0000-0001-7930-7977", "researcher": {"href": "https://publications.scilifelab.se/researcher/899ddd1ca8b148f696e5d30f30d762b1.json"}}, {"family": "Meineke", "given": "Birthe", "initials": "B", "orcid": "0000-0002-8912-0783", "researcher": {"href": "https://publications.scilifelab.se/researcher/e1cc783424be4fe2b41d82bba5c2a412.json"}}, {"family": "Bauer", "given": "Sebastian", "initials": "S", "orcid": "0009-0009-7265-1527", "researcher": {"href": "https://publications.scilifelab.se/researcher/b46eb06fbcd1476b8d55bd823a46a19c.json"}}, {"family": "Blom", "given": "Hans", "initials": "H", "orcid": "0000-0002-5584-9170", "researcher": {"href": "https://publications.scilifelab.se/researcher/3ce356a74dc84e0ea6af85397f11d869.json"}}, {"family": "Els\u00e4sser", "given": "Simon", "initials": "S", "orcid": "0000-0001-8724-4849", "researcher": {"href": "https://publications.scilifelab.se/researcher/fcf26e35e037499aa1441a7738ba61af.json"}}, {"family": "Brismar", "given": "Hjalmar", "initials": "H", "orcid": "0000-0003-0578-4003", "researcher": {"href": "https://publications.scilifelab.se/researcher/1ec23336e2ef4e298f340876f1136dce.json"}}], "type": "journal-article", "published": "2026-02-04", "journal": {"title": "Nano Lett.", "issn": "1530-6984", "volume": "26", "issue": "4", "pages": "1321-1326", "issn-l": null}, "abstract": "With recent advances in fluorescence microscopy, resolution is often limited by the size of the label and the resulting linkage error, rather than the microscope itself. Site-specific incorporation of noncanonical amino acids (ncAAs) combined with bioorthogonal click chemistry provides a powerful tool for fluorescent protein labeling, overcoming the spatial uncertainty inherent to antibody-based probes. Here, we present a method to further improve labeling precision by combining ncAA labeling with expansion microscopy (ExM) for dual-color super-resolution imaging. After optimizing labeling procedures and fluorophore selection, we visualize and resolve the nanoscale distribution of Na,K-ATPase \u03b11 and \u03b21 subunits in expanded HEK 293T cells. We validate our approach by super-resolution STED imaging of the ncAA labeled \u03b21 subunit in unexpanded cells. This work presents a strong framework for multiplexed, high-resolution imaging, suggesting that ncAA labeling combined with ExM enables biological imaging at the nanometer scale.", "doi": "10.1021/acs.nanolett.5c05301", "pmid": "41571281", "labels": {"Integrated Microscopy Technologies Stockholm": "Technology development"}, "xrefs": [{"db": "pmc", "key": "PMC12879918"}], "notes": [], "created": "2026-01-24T20:15:09.343Z", "modified": "2026-02-11T12:54:50.277Z"}