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Related Experiment Video

Updated: May 27, 2025

Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy

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Biocompatible Sulfonium-Based Covalent Probes For Endogenous Tubulin Fluorescence Nanoscopy In Live And Fixed Cells.

Gražvydas Lukinavičius¹, Marie Auvray¹, Tanja Koenen²

¹Max Planck Institute for Multidisciplinary Sciences.

Research Square

|February 20, 2025

Summary

Researchers developed a novel covalent fluorescent probe, 6-SiR-o-C-CTX, for precise tubulin imaging. This probe minimizes disruption to tubulin function and is compatible with super-resolution microscopy.

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Last Updated: May 27, 2025

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Area of Science:

Cell Biology
Biochemistry
Microscopy

Background:

Fluorescent probes are crucial for visualizing cellular dynamics, often enhanced by super-resolution imaging.
Conventional methods like fluorescent protein fusions or ligand-linked fluorophores can disrupt cellular functions.
Endogenous tubulin, vital for cell structure and division, requires precise imaging methods.

Purpose of the Study:

To develop a novel covalent fluorescent probe for endogenous tubulin.
To overcome limitations of existing probes, such as functional disruption and ligand effects.
To enable high-resolution imaging of tubulin with minimal impact on cellular processes.

Main Methods:

Design and synthesis of a novel covalent fluorescent probe, 6-SiR-o-C-CTX, based on cabazitaxel and silicon-rhodamine.
Incorporation of a biocompatible cleavable linker with a sulfonium center.
Validation in various human cell lines using confocal and STED nanoscopy.

Main Results:

The probe, 6-SiR-o-C-CTX, exhibited excellent cell permeability and fluorogenic properties.
Covalent labeling of endogenous tubulin was achieved across multiple human cell lines.
Targeting moiety washout preserved tubulin staining, minimizing functional disruption.
Compatibility with STED nanoscopy in live and fixed cells was demonstrated.

Conclusions:

6-SiR-o-C-CTX is an effective covalent fluorescent probe for high-resolution tubulin imaging.
The probe design minimizes disruption to tubulin function, addressing limitations of prior methods.
This technique offers a powerful tool for studying tubulin dynamics in cellular processes.