Digital-micromirror-device-based illumination strategies for background suppression in single-molecule localization...

Area of Science:

• Biophysics
• Optical Microscopy
• Super-resolution Imaging

Background:

• Single-molecule localization microscopy (SMLM) enables super-resolution imaging by precisely localizing fluorescent emitters.
• Background noise from out-of-focus light and autofluorescence degrades SMLM signal-to-noise ratio and localization precision.

Purpose of the Study:

• To develop novel illumination strategies for SMLM to improve imaging performance.
• To enhance the photon-to-background ratio and localization precision in SMLM.

Main Methods:

• Introduced two controllable illumination strategies utilizing a digital micromirror device (DMD).
• Implemented spatial and temporal modulation of illumination regions.
• Applied methods to SMLM acquisitions.

Main Results:

• Achieved significant enhancement of the photon-to-background ratio.
• Demonstrated improved localization precision in SMLM.
• Validated performance on challenging specimens like scattering tissues.

Conclusions:

• Controllable DMD-based illumination effectively boosts SMLM imaging performance.
• The proposed method enhances signal quality without added computational or instrumental complexity.
• Offers new possibilities for SMLM in difficult biological samples.