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Protein‐Specific, Multicolor and 3D STED Imaging in Cells with DNA‐Labeled Antibodies
Author(s) -
Spahn Christoph,
Hurter Florian,
Glaesmann Mathilda,
Karathanasis Christos,
Lampe Marko,
Heilemann Mike
Publication year - 2019
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201910115
Subject(s) - sted microscopy , photobleaching , fluorophore , confocal , confocal microscopy , microscopy , chemistry , oligonucleotide , fluorescence , fluorescence recovery after photobleaching , biophysics , fluorescence microscope , fluorescence lifetime imaging microscopy , dna , stimulated emission , microbiology and biotechnology , optics , biology , biochemistry , physics , laser
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.