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Time Multiplexing Super Resolving Technique for Imaging from a Moving Platform

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Published on: February 12, 2014

A multiemitter localization comparison of 3D superresolution imaging modalities.

Sheng Liu¹, Keith A Lidke

¹Department of Physics and Astronomy, University of New Mexico, Albuquerque, NM 87131 (USA).

Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry

|November 28, 2013

Summary

Superresolution imaging struggles with overlapping emitters, especially in 3D. This study found minimal performance differences between astigmatic, dual focal plane, and combined 3D imaging modalities for multi-emitter localization.

Keywords:

fluorescence microscopy fluorophores localization precision single-molecule studies superresolution imaging

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Area of Science:

Biophysics
Optical Microscopy
Nanotechnology

Background:

Single-molecule localization microscopy (SMLM) enables superresolution imaging.
Overlapping emission from multiple emitters complicates SMLM, particularly in 3D.
Localization precision in SMLM is affected by emitter overlap.

Purpose of the Study:

To compare the performance of different 3D single-molecule imaging modalities for localizing overlapping emitters.
To determine if specific 3D imaging techniques offer significant advantages for multi-emitter estimation.

Main Methods:

Investigated astigmatic imaging, dual focal plane imaging, and a combination of both.
Utilized the Cramér-Rao lower bound to quantify theoretical localization precision.
Evaluated performance across various conditions: photon counts, background noise, pixel size, and readout noise.

Main Results:

Differences in multi-emitter estimation performance between the compared 3D modalities were found to be small.
Localization precision is influenced by emitter overlap, but modality choice had minimal impact.

Conclusions:

Multiemitter fitting performance should not be the primary factor when selecting between astigmatic, dual focal plane, or combined 3D imaging modalities.
The choice of 3D imaging modality for SMLM does not significantly impact multi-emitter localization performance.