Open AccessBlinking Fluorescent Probes for Tubulin Nanoscopy in Living and Fixed Cells
Author(s) -
Ru Ta Gerasimaitė,
Jonas Bucevičius,
Kamila A. Kiszka,
Sebastian Schnorrenberg,
Georgij Kostiuk,
Tanja Koenen,
Gražvydas Lukinavičius
Publication year - 2021
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.1c00538
Subject(s) - microtubule , sted microscopy , tubulin , biophysics , fluorescence , linker , microscopy , fluorescence microscope , chemistry , biology , microbiology and biotechnology , physics , optics , stimulated emission , laser , computer science , operating system
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.