Reactivatable stimulated emission depletion microscopy using fluorescence-recoverable nanographene

Area of Science:

• Optics and Photonics
• Biophysics
• Materials Science

Background:

• Stimulated emission depletion (STED) microscopy achieves super-resolution imaging down to tens of nanometers.
• High depletion laser power improves resolution but causes irreversible fluorophore photo-bleaching, limiting imaging time.
• A trade-off exists between spatial resolution and imaging duration in conventional STED.

Purpose of the Study:

• To overcome the photo-bleaching limitation in STED microscopy.
• To introduce a novel imaging approach enabling prolonged high-resolution 3D STED imaging.
• To leverage the unique photophysical properties of nanographene dibenzo[hi,st]ovalene (DBOV) for STED applications.

Main Methods:

• Development of reactivatable STED (ReSTED) microscopy.
• Utilizing the nanographene dibenzo[hi,st]ovalene (DBOV) fluorophore.
• Employing near-infrared light for reversible fluorescence deactivation and reactivation.

Main Results:

• DBOV fluorescence is temporarily deactivated by STED light, not permanently photo-bleached.
• DBOV fluorescence can be reactivated by near-infrared light, including the 775 nm depletion beam.
• Hours-long, high-resolution 3D STED imaging was achieved using ReSTED with DBOV.

Conclusions:

• ReSTED microscopy with DBOV overcomes the photo-bleaching limitation of conventional STED.
• This approach enables extended durations for high-resolution 3D imaging.
• ReSTED significantly expands the applicability of STED microscopy in various scientific fields.