Ultrafast Super-Resolution Imaging Exploiting Spontaneous Blinking of Static Excimer Aggregates

Area of Science:

• Biophysics
• Nanotechnology
• Microscopy

Background:

• Single-molecule localization microscopy (SMLM) provides super-resolution imaging of biological structures.
• Slow imaging speeds due to limited fluorophore blinking frequency hinder SMLM applications.

Purpose of the Study:

• To develop an ultrafast SMLM technique for improved imaging speed.
• To overcome the limitations of conventional SMLM regarding fluorophore blinking frequency.

Main Methods:

• Exploited spontaneous fluorescence blinking of cyanine dye aggregates within DNA framework nanostructures.
• Utilized DNA templates to form static excimer aggregates acting as light-harvesting nanoantennas.
• Leveraged intermolecular excitation energy transfer (EET) for collective, ultrafast blinking of fluorophore aggregates.

Main Results:

• Achieved a 12.5-fold improvement in imaging speed for DNA nanostructures compared to conventional SMLM.
• Demonstrated real-time tracking of super-resolved DNA nanostructures for over 20 minutes in a microfluidic system.

Conclusions:

• The developed DNA framework-based strategy enables ultrafast SMLM.
• This technique significantly enhances imaging speed and holds potential for dynamic biomacromolecular studies in living cells.