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 Glogger, Christoph Spahn, Jörg Enderlein, Mike HeilemannORCiDGND
Parent Title (German):Angewandte Chemie
Place of publication:Weinheim
Document Type:Article
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
Page Number:4
First Page:6310
Last Page:6313
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.
Institutes:Biochemie, Chemie und Pharmazie
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften
Licence (German):License LogoCreative Commons - Namensnennung 4.0