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Journal Article

Rhodamine-Hoechst positional isomers for highly efficient staining of heterochromatin.

MPS-Authors
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Bucevicius,  J.
Laboratory of Chromatin Labeling and Imaging, Max Planck Institute for Biophysical Chemistry, Max Planck Society;

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Keller-Findeisen,  J.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

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Gilat,  T.
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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Lukinavicius,  G.
Laboratory of Chromatin Labeling and Imaging, Max Planck Institute for Biophysical Chemistry, Max Planck Society;

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Fulltext (public)

3029681.pdf
(Publisher version), 3MB

Supplementary Material (public)

3029681_Suppl_1.pdf
(Supplementary material), 10MB

3029681_Suppl_2.avi
(Supplementary material), 10MB

3029681_Suppl_3.avi
(Supplementary material), 942KB

Citation

Bucevicius, J., Keller-Findeisen, J., Gilat, T., Hell, S. W., & Lukinavicius, G. (2019). Rhodamine-Hoechst positional isomers for highly efficient staining of heterochromatin. Chemical Science, 10(7), 1962-1970. doi:10.1039/C8SC05082A.


Cite as: http://hdl.handle.net/21.11116/0000-0003-0DE8-A
Abstract
Hoechst conjugates to fluorescent dyes are popular DNA stains for live-cell imaging, but the relationship between their structure and performance remains elusive. This study of carboxyrhodamine–Hoechst 33258 conjugates reveals that a minimal change in the attachment point of the dye has dramatic effects on the properties of the final probe. All tested 6′-carboxyl dye-containing probes exhibited dual-mode binding to DNA and formed a dimmer complex at high DNA concentrations. The 5′-carboxyl dye-containing probes exhibited single-mode binding to DNA which translated into increased brightness and lower cytotoxicity. Up to 10-fold brighter nuclear staining by the newly developed probes allowed acquisition of stimulated emission depletion (STED) nanoscopy images of outstanding quality in living and fixed cells. Therefore we were able to estimate a diameter of ∼155 nm of the heterochromatin exclusion zones in the nuclear pore region in living cells and intact chicken erythrocytes and to localize telomeres relative to heterochromatin in living U-2 OS cells. Employing the highly efficient probes for two-color STED allowed visualization of DNA and tubulin structures in intact nucleated erythrocytes – a system where imaging is greatly hampered by high haemoglobin absorbance.