Multi‐Color, Bleaching‐Resistant Super‐Resolution Optical Fluctuation Imaging with Oligonucleotide‐Based Exchangeable Fluorophores
2021 | journal article; research paper. A publication with affiliation to the University of Göttingen.
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Multi‐Color, Bleaching‐Resistant Super‐Resolution Optical Fluctuation Imaging with Oligonucleotide‐Based Exchangeable Fluorophores
Glogger, M.; Spahn, C.; Enderlein, J. & Heilemann, M. (2021)
Angewandte Chemie International Edition, 60(12) pp. 6310-6313. DOI: https://doi.org/10.1002/anie.202013166
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- Authors
- Glogger, Marius; Spahn, Christoph; Enderlein, Jörg ; Heilemann, Mike
- Abstract
- Abstract 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.
Fluorophore labels that transiently bind to and unbind from a protein target allow photobleaching in super‐resolution optical fluctuation imaging (SOFI) to be circumvented. Using short fluorophore‐labeled DNA oligonucleotides, we demonstrate high‐contrast, low‐background, and multi‐color super‐resolution imaging with SOFI. image - Issue Date
- 2021
- Journal
- Angewandte Chemie International Edition
- Project
- EXC 2067: Multiscale Bioimaging
- Working Group
- RG Enderlein
- ISSN
- 1433-7851
- eISSN
- 1521-3773
- Language
- English
- Sponsor
- Deutsche Forschungsgemeinschaft http://dx.doi.org/10.13039/501100001659