Multi-color, bleaching-resistant super-resolution optical fluctuation imaging with oligonucleotide-based exchangeable fluorophores
- Super-resolution optical fluctuation imaging (SOFI) is a super-resolution microscopy technique that overcomes the diffraction limit by analyzing intensity fluctuations of statistically independent emitters in a time series of images. The final images are background-free and show confocality and enhanced spatial resolution (super-resolution). Fluorophore photobleaching, however, is a key limitation for recording long time series of images that will allow for the calculation of higher order SOFI results with correspondingly increased resolution. Here, we demonstrate that photobleaching can be circumvented by using fluorophore labels that reversibly and transiently bind to a target, and which are being replenished from a buffer which serves as a reservoir. Using fluorophore-labeled short DNA oligonucleotides, we labeled cellular structures with target-specific antibodies that contain complementary DNA sequences and record the fluctuation events caused by transient emitter binding. We show that this concept bypasses extensive photobleaching and facilitates two-color imaging of cellular structures with SOFI.
Author: | Marius GloggerORCiDGND, Christoph Klaus SpahnORCiDGND, Jörg Enderlein, Mike HeilemannORCiDGND |
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URN: | urn:nbn:de:hebis:30:3-638707 |
DOI: | https://doi.org/10.1002/anie.202013166 |
ISSN: | 1521-3773 |
Parent Title (German): | Angewandte Chemie |
Publisher: | Wiley-VCH |
Place of publication: | Weinheim |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2020/12/10 |
Date of first Publication: | 2020/12/10 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2022/03/16 |
Tag: | DNA-PAINT; SOFI; fluorescence; photobleaching; super-resolution microscopy |
Volume: | 60 |
Issue: | 12 |
Page Number: | 4 |
First Page: | 6310 |
Last Page: | 6313 |
Note: | M.G., C.S. and M.H. acknowledge funding by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation, Project-ID 259130777, SFB 1177). J.E. acknowledges support by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany's Excellence Strategy—EXC 2067/1-390729940. Open access funding enabled and organized by Projekt DEAL. |
HeBIS-PPN: | 493368922 |
Institutes: | Biochemie, Chemie und Pharmazie |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften |
Sammlungen: | Universitätspublikationen |
Licence (German): | ![]() |