Exchangeable fluorophore labels in super-resolution fluorescence microscopy

Abstract number
86
Presentation Form
Poster Flash Talk and Poster
DOI
10.22443/rms.elmi2021.86
Corresponding Email
[email protected]
Session
Poster Session 2
Authors
Marius Glogger (2), Christoph C. Spahn (3), Marko Lampe (1), Jörg Enderlein (4), Mike Heilemann (2)
Affiliations
1. Advanced Light Microscopy Facility, European Molecular Biology Laboratory, Meyerhofstr. 1, 69117 Heidelberg, Germany
2. Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt, 60438 Frankfurt, Germany
3. Max Planck Institute for Terrestrial Microbiology, 35043 Marburg, Germany
4. Third Institute of Physics - Biophysics, University of Göttingen, 37077 Göttingen, Germany
Keywords

Super-resolution microscopy, exchangeable dyes, SMLM

Abstract text

We present fluorophore labels that transiently and repetitively bind to their targets as probes for various types of super-resolution fluorescence microscopy. Transient labels typically show a weak affinity to a target, and exchange constantly with the buffer that constitutes a reservoir with a large amount of intact probes, leading to repetitive binding events to the same target (we refer to these labels as “exchangeable labels”). This dynamic labeling approach is insensitive to common photobleaching and yields a constant fluorescence signal over time, which has been successfully exploited in SMLM [1-3], STED [4, 5], single-particle tracking [6] and super-resolution optical fluctuation imaging (SOFI) [7]. We discuss properties of suitable exchangeable labels, experimental parameters for optimal performance for the different super-resolution methods, and present high-quality multicolor super-resolution imaging.

References

1.            Jungmann, R., et al., Single-molecule kinetics and super-resolution microscopy by fluorescence imaging of transient binding on DNA origami. Nano Lett, 2010. 10(11): p. 4756-61.

2.            Sharonov, A. and R.M. Hochstrasser, Wide-field subdiffraction imaging by accumulated binding of diffusing probes. Proc Natl Acad Sci U S A, 2006. 103(50): p. 18911-6.

3.            Spahn, C.K., et al., A toolbox for multiplexed super-resolution imaging of the E. coli nucleoid and membrane using novel PAINT labels. Sci Rep, 2018. 8(1): p. 14768.

4.            Spahn, C., et al., Whole-Cell, 3D, and Multicolor STED Imaging with Exchangeable Fluorophores. Nano Lett, 2019. 19(1): p. 500-505.

5.            Spahn, C., et al., Protein-Specific, Multicolor and 3D STED Imaging in Cells with DNA-Labeled Antibodies. Angew Chem Int Ed Engl, 2019. 58(52): p. 18835-18838.

6.            Harwardt, M.I.E., et al., Single-Molecule Super-Resolution Microscopy Reveals Heteromeric Complexes of MET and EGFR upon Ligand Activation. Int J Mol Sci, 2020. 21(8).

7.            Dertinger, T., et al., Fast, background-free, 3D super-resolution optical fluctuation imaging (SOFI). Proc Natl Acad Sci U S A, 2009. 106(52): p. 22287-92.