NanoPlex: a universal strategy for fluorescence microscopy multiplexing using nanobodies with erasable signals.

Autor: Mougios N; Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany.; Center for Biostructural Imaging of Neurodegeneration (BIN), University of Göttingen Medical Center, Göttingen, Germany., Cotroneo ER; Institute for Organic and Biomolecular Chemistry, University of Göttingen, Göttingen, Germany., Imse N; Institute for Organic and Biomolecular Chemistry, University of Göttingen, Göttingen, Germany., Setzke J; Center for Biostructural Imaging of Neurodegeneration (BIN), University of Göttingen Medical Center, Göttingen, Germany., Rizzoli SO; Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), University of Göttingen, Göttingen, Germany., Simeth NA; Institute for Organic and Biomolecular Chemistry, University of Göttingen, Göttingen, Germany.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), University of Göttingen, Göttingen, Germany., Tsukanov R; III. Institute of Physics - Biophysics, Georg August University, Göttingen, Germany., Opazo F; Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany. fopazo@gwdg.de.; Center for Biostructural Imaging of Neurodegeneration (BIN), University of Göttingen Medical Center, Göttingen, Germany. fopazo@gwdg.de.; NanoTag Biotechnologies GmbH, Göttingen, Germany. fopazo@gwdg.de.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 Oct 10; Vol. 15 (1), pp. 8771. Date of Electronic Publication: 2024 Oct 10.
DOI: 10.1038/s41467-024-53030-w
Abstrakt: Fluorescence microscopy has long been a transformative technique in biological sciences. Nevertheless, most implementations are limited to a few targets, which have been revealed using primary antibodies and fluorescently conjugated secondary antibodies. Super-resolution techniques such as Exchange-PAINT and, more recently, SUM-PAINT have increased multiplexing capabilities, but they require specialized equipment, software, and knowledge. To enable multiplexing for any imaging technique in any laboratory, we developed NanoPlex, a streamlined method based on conventional antibodies revealed by engineered secondary nanobodies that allow the selective removal of fluorescence signals. We develop three complementary signal removal strategies: OptoPlex (light-induced), EnzyPlex (enzymatic), and ChemiPlex (chemical). We showcase NanoPlex reaching 21 targets for 3D confocal analyses and 5-8 targets for dSTORM and STED super-resolution imaging. NanoPlex has the potential to revolutionize multi-target fluorescent imaging methods, potentially redefining the multiplexing capabilities of antibody-based assays.
(© 2024. The Author(s).)
Databáze: MEDLINE
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