Blinking Fluorescent Probes for Tubulin Nanoscopy in Living and Fixed Cells

• DOI: 10.1021/acschembio.1c00538
• Publication Date: 2021-11-04T15:34:25Z
• Authors: Ru̅ta Gerasimaitė; Jonas Bucevičius; Kamila A. Kiszka; Sebastian Schnorr...; Georgij Kostiuk; Tanja Koenen; Gražvydas Lukinav...
• Keywords: STED microscopy; Linker

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