Protein‐Specific, Multicolor and 3D STED Imaging in Cells with DNA‐Labeled Antibodies. Issue 52 (7th November 2019)

• Record Type: Journal Article
• Authors: Spahn, Christoph; Hurter, Florian; Glaesmann, Mathilda; Karathanasis, Christos; Lampe, Marko; Heilemann, Mike
• Is Part Of: Angewandte Chemie international edition. Volume 58:Issue 52(2019)
• Page Start: 18835
• Page End: 18838
• Publication Date: 2019-11-07
• Subjects: DNA-PAINT -- fluorescence -- fluorescent probes -- multicolor imaging -- STED microscopy; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201910115
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: 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. Abstract : Short, DNA‐based fluorophore labels that transiently and repeatedly bind to DNA‐labeled antibodies enable super‐resolution STED imaging. Compared to covalent labels, dynamic labels constantly exchange between target‐bound and free states, which bypasses photobleaching and subsequently increases image quality and information content.