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Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging

Time-gated Förster resonance energy transfer (FRET) using the unique material combination of long-lifetime terbium complexes (Tb) and semiconductor quantum dots (QDs) provides many advantages for highly sensitive and multiplexed biosensing. Although time-gated detection can efficiently suppress samp...

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Autores principales: Afsari, Hamid Samareh, Cardoso Dos Santos, Marcelina, Lindén, Stina, Chen, Ting, Qiu, Xue, van Bergen en Henegouwen, Paul M. P., Jennings, Travis L., Susumu, Kimihiro, Medintz, Igor L., Hildebrandt, Niko, Miller, Lawrence W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4928903/
https://www.ncbi.nlm.nih.gov/pubmed/27386579
http://dx.doi.org/10.1126/sciadv.1600265
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author Afsari, Hamid Samareh
Cardoso Dos Santos, Marcelina
Lindén, Stina
Chen, Ting
Qiu, Xue
van Bergen en Henegouwen, Paul M. P.
Jennings, Travis L.
Susumu, Kimihiro
Medintz, Igor L.
Hildebrandt, Niko
Miller, Lawrence W.
author_facet Afsari, Hamid Samareh
Cardoso Dos Santos, Marcelina
Lindén, Stina
Chen, Ting
Qiu, Xue
van Bergen en Henegouwen, Paul M. P.
Jennings, Travis L.
Susumu, Kimihiro
Medintz, Igor L.
Hildebrandt, Niko
Miller, Lawrence W.
author_sort Afsari, Hamid Samareh
collection PubMed
description Time-gated Förster resonance energy transfer (FRET) using the unique material combination of long-lifetime terbium complexes (Tb) and semiconductor quantum dots (QDs) provides many advantages for highly sensitive and multiplexed biosensing. Although time-gated detection can efficiently suppress sample autofluorescence and background fluorescence from directly excited FRET acceptors, Tb-to-QD FRET has rarely been exploited for biomolecular imaging. We demonstrate Tb-to-QD time-gated FRET nanoassemblies that can be applied for intra- and extracellular imaging. Immunostaining of different epitopes of the epidermal growth factor receptor (EGFR) with Tb- and QD-conjugated antibodies and nanobodies allowed for efficient Tb-to-QD FRET on A431 cell membranes. The broad usability of Tb-to-QD FRET was further demonstrated by intracellular Tb-to-QD FRET and Tb-to-QD-to-dye FRET using microinjection as well as cell-penetrating peptide–mediated endocytosis with HeLa cells. Effective brightness enhancement by FRET from several Tb to the same QD, the use of low nanomolar concentrations, and the quick and sensitive detection void of FRET acceptor background fluorescence are important advantages for advanced intra- and extracellular imaging of biomolecular interactions.
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spelling pubmed-49289032016-07-06 Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging Afsari, Hamid Samareh Cardoso Dos Santos, Marcelina Lindén, Stina Chen, Ting Qiu, Xue van Bergen en Henegouwen, Paul M. P. Jennings, Travis L. Susumu, Kimihiro Medintz, Igor L. Hildebrandt, Niko Miller, Lawrence W. Sci Adv Research Articles Time-gated Förster resonance energy transfer (FRET) using the unique material combination of long-lifetime terbium complexes (Tb) and semiconductor quantum dots (QDs) provides many advantages for highly sensitive and multiplexed biosensing. Although time-gated detection can efficiently suppress sample autofluorescence and background fluorescence from directly excited FRET acceptors, Tb-to-QD FRET has rarely been exploited for biomolecular imaging. We demonstrate Tb-to-QD time-gated FRET nanoassemblies that can be applied for intra- and extracellular imaging. Immunostaining of different epitopes of the epidermal growth factor receptor (EGFR) with Tb- and QD-conjugated antibodies and nanobodies allowed for efficient Tb-to-QD FRET on A431 cell membranes. The broad usability of Tb-to-QD FRET was further demonstrated by intracellular Tb-to-QD FRET and Tb-to-QD-to-dye FRET using microinjection as well as cell-penetrating peptide–mediated endocytosis with HeLa cells. Effective brightness enhancement by FRET from several Tb to the same QD, the use of low nanomolar concentrations, and the quick and sensitive detection void of FRET acceptor background fluorescence are important advantages for advanced intra- and extracellular imaging of biomolecular interactions. American Association for the Advancement of Science 2016-06-10 /pmc/articles/PMC4928903/ /pubmed/27386579 http://dx.doi.org/10.1126/sciadv.1600265 Text en Copyright © 2016, The Authors http://creativecommons.org/licenses/by-nc/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (http://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.
spellingShingle Research Articles
Afsari, Hamid Samareh
Cardoso Dos Santos, Marcelina
Lindén, Stina
Chen, Ting
Qiu, Xue
van Bergen en Henegouwen, Paul M. P.
Jennings, Travis L.
Susumu, Kimihiro
Medintz, Igor L.
Hildebrandt, Niko
Miller, Lawrence W.
Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging
title Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging
title_full Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging
title_fullStr Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging
title_full_unstemmed Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging
title_short Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging
title_sort time-gated fret nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4928903/
https://www.ncbi.nlm.nih.gov/pubmed/27386579
http://dx.doi.org/10.1126/sciadv.1600265
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