Protein-Specific, Multicolor and 3D STED Imaging in Cells with DNA-Labeled Antibodies.
DNA-PAINT
STED microscopy
fluorescence
fluorescent probes
multicolor imaging
Journal
Angewandte Chemie (International ed. in English)
ISSN: 1521-3773
Titre abrégé: Angew Chem Int Ed Engl
Pays: Germany
ID NLM: 0370543
Informations de publication
Date de publication:
19 12 2019
19 12 2019
Historique:
received:
08
08
2019
revised:
25
09
2019
pubmed:
12
10
2019
medline:
25
9
2020
entrez:
12
10
2019
Statut:
ppublish
Résumé
Photobleaching is a major challenge in fluorescence microscopy, in particular if high excitation light intensities are used. Signal-to-noise and spatial resolution may be compromised, which limits the amount of information that can be extracted from an image. Photobleaching can be bypassed by using exchangeable labels, which transiently bind to and dissociate from a target, thereby replenishing the destroyed labels with intact ones from a reservoir. Here, we demonstrate confocal and STED microscopy with short, fluorophore-labeled oligonucleotides that transiently bind to complementary oligonucleotides attached to protein-specific antibodies. The constant exchange of fluorophore labels in DNA-based STED imaging bypasses photobleaching that occurs with covalent labels. We show that this concept is suitable for targeted, two-color STED imaging of whole cells.
Identifiants
pubmed: 31603612
doi: 10.1002/anie.201910115
pmc: PMC6972974
doi:
Substances chimiques
Antibodies
0
Proteins
0
DNA
9007-49-2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
18835-18838Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : HE 6166/11-1
Pays : International
Organisme : Deutsche Forschungsgemeinschaft
ID : HE 6166/17-1
Pays : International
Organisme : Deutsche Forschungsgemeinschaft
ID : SFB 902
Pays : International
Organisme : Deutsche Forschungsgemeinschaft
ID : SFB 807
Pays : International
Organisme : Deutsche Forschungsgemeinschaft
ID : SFB 1177
Pays : International
Informations de copyright
© 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.
Références
Angew Chem Int Ed Engl. 2015 Aug 24;54(35):10216-9
pubmed: 26201868
Q Rev Biophys. 2015 May;48(2):178-243
pubmed: 25998828
Angew Chem Int Ed Engl. 2017 Mar 27;56(14):4052-4055
pubmed: 28256790
Opt Express. 2015 Apr 6;23(7):8630-8
pubmed: 25968701
Nat Methods. 2013 Jun;10(6):557-62
pubmed: 23624665
Proc Natl Acad Sci U S A. 2017 Sep 12;114(37):9797-9802
pubmed: 28847959
Chem Sci. 2017 Oct 1;8(10):7138-7142
pubmed: 29147545
Nano Lett. 2019 Jan 9;19(1):500-505
pubmed: 30525682
Chem Rev. 2017 Jun 14;117(11):7478-7509
pubmed: 28287710
Nano Lett. 2010 Nov 10;10(11):4756-61
pubmed: 20957983
Angew Chem Int Ed Engl. 2019 Dec 19;58(52):18835-18838
pubmed: 31603612
Chem Rev. 2017 Jun 14;117(11):7377-7427
pubmed: 28262022
Angew Chem Int Ed Engl. 2019 Sep 9;58(37):13004-13008
pubmed: 31314157
Sci Rep. 2018 Oct 3;8(1):14768
pubmed: 30282984
Nat Commun. 2016 Jan 29;7:10372
pubmed: 26822409
Nat Protoc. 2017 Jun;12(6):1198-1228
pubmed: 28518172
Nat Methods. 2018 Sep;15(9):685-688
pubmed: 30127504
Curr Opin Chem Biol. 2017 Aug;39:32-38
pubmed: 28544971
J Struct Biol. 2013 Sep;183(3):363-367
pubmed: 23684965