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Long-Term Super-Resolution Imaging through Torsional Angle Inversed Rhodamines
Xue Zhang1, Ying Zheng1,2, Lujia Yang1
1State Key Laboratory of Fine Chemicals, Frontiers Science Center for Smart Materials Oriented Chemical Engineering, Dalian University of Technology, Dalian 116024, China.
Analytical Chemistry
|October 22, 2025
Summary
Researchers developed a novel crowding strategy to enhance super-resolution imaging by engineering silicon-substituted rhodamine dyes. This method prolongs imaging duration, enabling detailed observation of cellular structures in living cells.
Area of Science:
- Biophysics
- Chemical Biology
- Cell Biology
Background:
- Super-resolution microscopy requires stable fluorophores for detailed cellular imaging.
- Fluorophore blinking limitations obscure dynamic protein arrangements within cells.
Purpose of the Study:
- To develop a strategy for prolonging fluorophore blinking events in super-resolution imaging.
- To engineer novel rhodamine derivatives with enhanced stability for live-cell imaging.
Main Methods:
- Engineered sulfonamide rhodamines via atom-radii expansion (O-C-Si) to create steric repulsion.
- Investigated geometric electron effects from bridging atom substitution.
- Analyzed single-molecule kinetics, including recruiting rates and survival lifetimes.
Main Results:
- Developed silicon-substituted rhodamines exhibiting decreased self-blinking kinetics.
- Demonstrated prolonged single-molecule survival lifetimes and reduced recruiting rates.
- Achieved persistent molecular localization imaging of suborganelle proteins for up to 0.5 hours in living cells.
Conclusions:
- The hybrid crowding strategy effectively prolongs imaging time in super-resolution microscopy.
- Silicon-substituted rhodamines offer versatile capabilities for 3D and dual-color live-cell imaging.
- Structural engineering of fluorophores is a promising approach to overcome blinking limitations in imaging.
