Theoretical analysis of spectral precision in spectroscopic single-molecule localization microscopy

Area of Science:

• Optical Microscopy
• Spectroscopy
• Nanotechnology

Background:

• Spectroscopic single-molecule localization microscopy (sSMLM) offers simultaneous nanoscopic localization and spectral information.
• Establishing a theoretical framework for spectral precision is crucial for sSMLM system optimization.

Purpose of the Study:

• To develop a theoretical model for analyzing spectral precision in sSMLM.
• To guide the design and optimization of sSMLM systems by understanding key parameters.

Main Methods:

• Numerical simulations and analytical solutions were employed to create the theoretical model.
• The model incorporates signal and background shot noises, readout noise, and spectral calibration.

Main Results:

• A delicate balance among system parameters is essential for optimal spectral precision.
• The highest spectral precision is achieved at a specific system spectral dispersion.
• For instance, 3000 photons and 2 e- readout noise require 1.6 nm/pixel dispersion for 1.31 nm precision.

Conclusions:

• The theoretical model provides a foundation for understanding and improving sSMLM spectral precision.
• System spectral dispersion is a critical factor in achieving optimal performance in sSMLM imaging.