English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Combined use of unnatural amino acids enables dual color super-resolution Imaging of proteins via click chemistry.

MPS-Authors
There are no MPG-Authors available
External Ressource
No external resources are shared
Fulltext (public)
There are no public fulltexts stored in PuRe
Supplementary Material (public)

3015436_Suppl_1.pdf
(Supplementary material), 2MB

3015436_Suppl_2.pdf
(Supplementary material), 23MB

Citation

Saal, K. A., Richter, F., Rehling, P., & Rizzoli, S. O. (2018). Combined use of unnatural amino acids enables dual color super-resolution Imaging of proteins via click chemistry. ACS Nano, 12(12), 12247-12254. doi:10.1021/acsnano.8b06047.


Cite as: http://hdl.handle.net/21.11116/0000-0002-AF7F-C
Abstract
Recent advances in optical nanoscopy have brought the imaging resolution to the size of the individual macromolecules, thereby setting stringent requirements for the fluorescent labels. Such requirements are optimally fulfilled by the incorporation of unnatural amino acids (UAAs) in the proteins of interest (POI), followed by fluorophore conjugation via click chemistry. However, this approach has been limited to single POIs in mammalian cells. Here we solve this problem by incorporating different UAAs in different POIs, which are expressed in independent cell sets. The cells are then fused, thereby combining the different proteins and organelles, and are easily imaged by dual-color super-resolution microscopy. This procedure, which we termed Fuse2Click, is simple, requires only the well-established Amber codon, and allows the use of all previously optimized UAAs and tRNA/RS pairs. This should render it a tool of choice for multi-color click-based imaging.