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  DNA-barcoded labeling probes for highly multiplexed Exchange-PAINT imaging

Agasti, S. S., Wang, Y., Schueder, F., Sukumar, A., Jungmann, R., & Yin, P. (2017). DNA-barcoded labeling probes for highly multiplexed Exchange-PAINT imaging. Chemical Science, 8(4), 3080-3091. doi:10.1039/c6sc05420j.

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 Urheber:
Agasti, Sarit S.1, Autor
Wang, Yu1, Autor
Schueder, Florian2, Autor           
Sukumar, Aishwarya1, Autor
Jungmann, Ralf2, Autor           
Yin, Peng1, Autor
Affiliations:
1external, ou_persistent22              
2Jungmann, Ralf / Molecular Imaging and Bionanotechnology, Max Planck Institute of Biochemistry, Max Planck Society, ou_2149679              

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Schlagwörter: SUPERRESOLUTION MICROSCOPY; FLUORESCENT-PROBES; RESOLUTION LIMIT; NANOBODIES; BINDING; ACTIN; TECHNOLOGY; FILAMENT; CELLS; STORMChemistry;
 Zusammenfassung: Recent advances in super-resolution fluorescence imaging allow researchers to overcome the classical diffraction limit of light, and are already starting to make an impact in biology. However, a key challenge for traditional super-resolution methods is their limited multiplexing capability, which prevents a systematic understanding of multi-protein interactions on the nanoscale. Exchange-PAINT, a recently developed DNA-based multiplexing approach, in theory facilitates spectrally-unlimited multiplexing by sequentially imaging target molecules using orthogonal dye-labeled 'imager' strands. While this approach holds great promise for the bioimaging community, its widespread application has been hampered by the availability of DNA-conjugated ligands for protein labeling. Herein, we report a universal approach for the creation of DNA-barcoded labeling probes for highly multiplexed Exchange-PAINT imaging, using a variety of affinity reagents such as primary and secondary antibodies, nanobodies, and small molecule binders. Furthermore, we extend the availability of orthogonal imager strands for Exchange-PAINT to over 50 and assay their orthogonality in a novel DNA origami-based crosstalk assay. Using our optimized conjugation and labeling strategies, we demonstrate nine-color super-resolution imaging in situ in fixed cells.

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Sprache(n): eng - English
 Datum: 2017-01-302017
 Publikationsstatus: Erschienen
 Seiten: 12
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: -
 Identifikatoren: ISI: 000397560500072
DOI: 10.1039/c6sc05420j
 Art des Abschluß: -

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Projektname : MolMap, Grant agreement number 680241
Grant ID : 680241
Förderprogramm : ERC Starting Grant
Förderorganisation : -

Quelle 1

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Titel: Chemical Science
  Andere : Chem. Sci.
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: Cambridge, UK : Royal Society of Chemistry
Seiten: - Band / Heft: 8 (4) Artikelnummer: - Start- / Endseite: 3080 - 3091 Identifikator: ISSN: 2041-6520
CoNE: https://pure.mpg.de/cone/journals/resource/2041-6520