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  Blinking fluorescent probes for tubulin nanoscopy in living and fixed cells

Gerasimaite, R., Bucevicius, J., Kiszka, K., Schnorrenberg, S., Kostiuk, G., Koenen, T., et al. (2021). Blinking fluorescent probes for tubulin nanoscopy in living and fixed cells. ACS Chemical Biology, 16(11), 2130-2136. doi:10.1021/acschembio.1c00538.

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Gerasimaite, R.1, Author              
Bucevicius, J.1, Author              
Kiszka, K.2, Author              
Schnorrenberg, S., Author
Kostiuk, G.1, Author              
Koenen, T.2, Author              
Lukinavičius, G.1, Author              
Affiliations:
1Laboratory of Chromatin Labeling and Imaging, Max Planck Institute for Biophysical Chemistry, Max Planck Society, ou_2616691              
2Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society, Göttingen, DE, ou_578627              

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 Abstract: Here we report a small molecule tubulin probe for single-molecule localization microscopy (SMLM), stimulated emission depletion (STED) microscopy and MINFLUX nanoscopy, which can be used in living and fixed cells. We explored a series of taxane derivatives containing spontaneously blinking far-red dye hydroxymethyl silicon–rhodamine (HMSiR) and found that the linker length profoundly affects the probe permeability and off-targeting in living cells. The best performing probe, HMSiR-tubulin, is composed of cabazitaxel and the 6′-regioisomer of HMSiR bridged by a C6 linker. Microtubule diameter of ≤50 nm was routinely measured in SMLM experiments on living and fixed cells. HMSiR-tubulin allows a complementary use of different nanoscopy techniques for investigating microtubule functions and developing imaging methods. For the first time, we resolved the inner microtubule diameter of 16 ± 5 nm by optical nanoscopy and thereby demonstrated the utility of a self-blinking dye for MINFLUX imaging.

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Language(s): eng - English
 Dates: 2021-11-042021-11-19
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1021/acschembio.1c00538
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Title: ACS Chemical Biology
Source Genre: Journal
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Pages: - Volume / Issue: 16 (11) Sequence Number: - Start / End Page: 2130 - 2136 Identifier: ISSN: 1554-8929
ISSN: 1554-8937