Precisely and accurately localizing single emitters in fluorescence microscopy

Area of Science:

• Optics and Photonics
• Biophysics
• Nanotechnology

Background:

• Single-molecule localization and photophysics enable nanoscale imaging with visible light.
• These methods are crucial for single-particle tracking (SPT) and super-resolution microscopy.
• Localization precision and accuracy are critical for optimizing these techniques.

Purpose of the Study:

• To synthesize developments in single-fluorophore and nanoparticle localization precision and accuracy.
• To discuss the practical implications of these advancements.
• To guide researchers in SPT and super-resolution localization microscopy.

Main Methods:

• Review of existing literature on single-molecule localization techniques.
• Analysis of photophysical principles influencing localization.
• Synthesis of practical considerations for achieving high precision and accuracy.

Main Results:

• Detailed overview of factors affecting localization precision and accuracy.
• Discussion of trade-offs between different localization algorithms and hardware.
• Highlighting methods to improve localization performance.

Conclusions:

• Advancements in localization precision and accuracy are essential for progress in nanoscale imaging.
• Understanding these developments empowers researchers to enhance their single-particle tracking and super-resolution microscopy experiments.
• This synthesis provides a valuable resource for the scientific community.