English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Multicolour nanoscopy of fixed and living cells with a single STED beam and hyperspectral detection.

MPS-Authors
/persons/resource/persons205119

Winter,  F.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons205123

Loidolt,  M.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons16012

Westphal,  V.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons191595

Butkevich,  A.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons185783

Gregor,  C.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons15739

Sahl,  S. J.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons15210

Hell,  S. W.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

External Ressource
No external resources are shared
Fulltext (public)

2430484.pdf
(Publisher version), 3MB

Supplementary Material (public)

2430484_Suppl.pdf
(Supplementary material), 3MB

Citation

Winter, F., Loidolt, M., Westphal, V., Butkevich, A., Gregor, C., Sahl, S. J., et al. (2017). Multicolour nanoscopy of fixed and living cells with a single STED beam and hyperspectral detection. Scientific Reports, 7: 46492. doi:10.1038/srep46492.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002D-29CB-A
Abstract
The extension of fluorescence nanoscopy to larger numbers of molecular species concurrently visualized by distinct markers is of great importance for advanced biological applications. To date, up to four markers had been distinguished in STED experiments featuring comparatively elaborate imaging schemes and optical setups, and exploiting various properties of the fluorophores. Here we present a simple yet versatile STED design for multicolour imaging below the diffraction limit. A hyperspectral detection arrangement (hyperSTED) collects the fluorescence in four spectral channels, allowing the separation of four markers with only one excitation wavelength and a single STED beam. Unmixing of the different marker signals based on the simultaneous readout of all channels is performed with a non-negative matrix factorization algorithm. We illustrate the approach showing four-colour nanoscopy of fixed and living cellular samples.