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Updated: Feb 25, 2026
Whole-cell Super-Resolution Imaging via DNA-PAINT on a Spinning Disk Confocal with Optical Photon Reassignment
Published on: January 6, 2026
Reactive oxygen species-resistant ultrastable super-resolution DNA framework dots.
Chengpin Liang1, Qingting Li1,2, Bin Chen1
1State Key Laboratory of Synergistic Chem-Bio Synthesis, School of Chemistry and Chemical Engineering, New Cornerstone Science Laboratory, Frontiers Science Center for Transformative Molecules and National Center for Translational Medicine, Shanghai Jiao Tong University, Shanghai 200240, China.
Researchers developed photobleaching-resistant DNA framework (SDF) dots that mimic the natural green fluorescent protein (GFP). These ultrastable SDF dots enhance fluorophore photostability for advanced super-resolution imaging applications.
Area of Science:
- Biophysics and biomedical research
- Materials science
- Nanotechnology
Background:
- Nanoconfinement in natural and artificial systems modulates molecular properties for diverse applications.
- Green fluorescent protein (GFP) utilizes a peptide scaffold for chromophore stabilization via confinement.
- Developing artificial systems for controlled molecular confinement is crucial for advanced optical applications.
Purpose of the Study:
- To engineer photobleaching-resistant super-resolution DNA framework (SDF) dots for programmable fluorophore confinement, inspired by GFP.
- To enhance fluorophore photostability and enable advanced super-resolution microscopy techniques.
- To create a versatile platform for developing ultrastable fluorescent probes.
Main Methods:
- Development of DNA framework (SDF) dots with an internal cavity for fluorophore encapsulation.
- Assessment of photobleaching resistance against reactive oxygen species.
- Quantification of photostability enhancement with varying fluorophore labeling strategies within SDF dots.
- Application of SDF dots in super-resolution imaging techniques like stimulated emission depletion (STED) and structured illumination microscopy (SIM).
Main Results:
- SDF dots exhibit significant resistance to reactive oxygen species-induced photobleaching due to DNA framework shielding.
- Fluorophore encapsulation within SDF dots resulted in up to ~1.8-fold and ~50-fold enhancement in photostability compared to corner labeling and single labeling on DNA, respectively.
- Successful STED imaging of live cell membranes for over 30 minutes was achieved.
- Development of ultrastable super-resolution SIM barcodes capable of distinguishing eighteen colors with ~70 nm spatial resolution.
Conclusions:
- SDF dots provide a robust and adaptable platform for engineering ultrastable fluorescent probes.
- The developed nanoconfinement strategy significantly enhances fluorophore photostability for super-resolution imaging.
- This approach offers a versatile tool for advancing single-particle tracking and imaging in biophysics and biomedical research.
