期刊
ACS CHEMICAL BIOLOGY
卷 16, 期 11, 页码 2130-2136出版社
AMER CHEMICAL SOC
DOI: 10.1021/acschembio.1c00538
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资金
- Max Planck Society
A small molecule tubulin probe has been developed for use in single-molecule localization microscopy, STED microscopy, and MINFLUX nanoscopy, enabling measurement of microtubule diameter in living and fixed cells. The probe, HMSiR-tubulin, allows for complementary use of different nanoscopy techniques and has demonstrated utility in resolving inner microtubule diameter for the first time.
Here we report a small molecule tubulin probe for single-molecule localization microscopy (SMLM), stimulated emission depletion (STED) microscopy and MINFLUX nano-scopy, which can be used in living and fixed cells. We explored a series of taxane derivatives containing spontaneously blinking farred 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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