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Two-color 810 nm STED nanoscopy of living cells with endogenous SNAP-tagged fusion proteins.

MPS-Authors
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Butkevich,  A.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Ta,  H.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Ratz,  M.
Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society;

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Stoldt,  S.
Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society;

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Jakobs,  S.
Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society;

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Belov,  V. N.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Hell,  S. W.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Supplementary Material (public)

2483822_Suppl.pdf
(Supplementary material), 4MB

Citation

Butkevich, A., Ta, H., Ratz, M., Stoldt, S., Jakobs, S., Belov, V. N., et al. (2018). Two-color 810 nm STED nanoscopy of living cells with endogenous SNAP-tagged fusion proteins. ACS Chemical Biology, 13(2), 475-480. doi:10.1021/acschembio.7b00616.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002D-FCD8-A
Abstract
A 810 nm STED nanoscopy setup and an appropriate combination of two fluorescent dyes (Si-rhodamine 680SiR and carbopyronine 610CP) have been developed for near-IR live-cell superresolution imaging. Vimentin endogenously tagged using the CRISPR/Cas9 approach with the SNAP-tag, together with a non-covalent tubulin label, provided reliable and cell-to-cell reproducible dual-color confocal and STED imaging of the cytoskeleton in living cells.