Protein-specific, multicolor and 3D STED imaging in cells with DNA-labeled antibodies
- 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.
Download full text files
Additional Services
| Author: | Christoph Klaus SpahnORCiDGND, Florian Hurter, Mathilda Glaesmann, Christos KarathanasisORCiDGND, Marko LampeORCiDGND, Mike HeilemannORCiDGND |
|---|---|
| URN: | urn:nbn:de:hebis:30:3-638176 |
| DOI: | https://doi.org/10.1002/anie.201910115 |
| ISSN: | 1521-3773 |
| Parent Title (English): | Angewandte Chemie |
| Publisher: | Wiley-VCH |
| Place of publication: | Weinheim |
| Document Type: | Article |
| Language: | English |
| Date of Publication (online): | 2019/10/11 |
| Date of first Publication: | 2019/10/11 |
| Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
| Release Date: | 2022/01/25 |
| Tag: | DNA-PAINT; STED microscopy; fluorescence; fluorescent probes; multicolor imaging |
| Volume: | 58 |
| Issue: | 52 |
| Page Number: | 4 |
| First Page: | 18835 |
| Last Page: | 18838 |
| Note: | M.H., M.G., C.K., and C.S. acknowledge funding by the German Science Foundation (SFB 902, SFB 807, HE6166/17-1, HE6166/11-1). |
| HeBIS-PPN: | 490966837 |
| Institutes: | Biochemie, Chemie und Pharmazie |
| Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften |
| 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie | |
| Sammlungen: | Universitätspublikationen |
| Licence (German): |  Creative Commons - Namensnennung 4.0 |
