Time-correlated single molecule localization microscopy enhances resolution and fidelity

Area of Science:

• Biophysics
• Optical Microscopy
• Superresolution Imaging

Background:

• Single-molecule-localization-microscopy (SMLM) achieves nanoscale resolution but often ignores emitter intensity trajectory data.
• Existing SMLM reconstruction methods have limitations in maximizing spatial resolution and fidelity.

Purpose of the Study:

• To introduce time-correlated-SMLM (tcSMLM) for enhanced SMLM reconstruction.
• To leverage the full intensity trajectory information of individual emitters for improved imaging.
• To provide guidelines for applying tcSMLM to various SMLM datasets.

Main Methods:

• Development of the time-correlated-SMLM (tcSMLM) approach.
• Utilizing complete intensity trajectories of single emitters in reconstruction.
• Testing tcSMLM with simulated and experimental SMLM data.

Main Results:

• tcSMLM significantly increases spatial resolution and reconstruction fidelity.
• Improvements are particularly notable under low signal-to-noise ratio (SNR), high acquisition rates, and high emitter densities.
• The method is compatible with existing SMLM and superresolution algorithms.

Conclusions:

• tcSMLM effectively enhances SMLM imaging by incorporating temporal intensity information.
• The approach offers improved performance in demanding imaging scenarios.
• tcSMLM is a versatile and accessible tool for advancing superresolution microscopy.