English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Exchangeable HaloTag ligands for super-resolution fluorescence microscopy

MPS-Authors
/persons/resource/persons281819

Kompa,  Julian
Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society;

/persons/resource/persons286136

Bruins,  Jorick
Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society;

/persons/resource/persons264410

Wilhelm,  Jonas
Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society;

/persons/resource/persons228818

Frei,  Michelle S.
Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society;

/persons/resource/persons141821

D´Este,  Elisa
Max Planck Institute for Medical Research, Max Planck Society;

/persons/resource/persons212621

Hiblot,  Julien
Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society;

/persons/resource/persons203696

Johnsson,  Kai
Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society;

Fulltext (restricted access)
There are currently no full texts shared for your IP range.
Fulltext (public)
There are no public fulltexts stored in PuRe
Supplementary Material (public)
There is no public supplementary material available
Citation

Kompa, J., Bruins, J., Glogger, M., Wilhelm, J., Frei, M. S., Tarnawski, M., et al. (2022). Exchangeable HaloTag ligands for super-resolution fluorescence microscopy. Journal of the American Chemical Society, 145(5), 3075-3083. doi:10.1021/jacs.2c11969.


Cite as: https://hdl.handle.net/21.11116/0000-000C-83A0-A
Abstract
The specific and covalent labeling of the protein HaloTag with fluorescent probes in living cells makes it a powerful tool for bioimaging. However, the irreversible attachment of the probe to HaloTag precludes imaging applications that require transient binding of the probe and comes with the risk of irreversible photobleaching. Here, we introduce exchangeable ligands for fluorescence labeling of HaloTag (xHTLs) that reversibly bind to HaloTag and that can be coupled to rhodamines of different colors. In stimulated emission depletion (STED) microscopy, probe exchange of xHTLs allows imaging with reduced photobleaching as compared to covalent HaloTag labeling. Transient binding of fluorogenic xHTLs to HaloTag fusion proteins enables points accumulation for imaging in nanoscale topography (PAINT) and MINFLUX microscopy. We furthermore introduce pairs of xHTLs and HaloTag mutants for dual-color PAINT and STED microscopy. xHTLs thus open up new possibilities in imaging across microscopy platforms for a widely used labeling approach.