Information-Efficient, Off-Center Sampling Results in Improved Precision in 3D Single Particle Tracking Microscopy

2021 ◽  
Author(s):  
Kevin Welsher ◽  
Chen Zhang

In this work, we present a 3D single-particle tracking system that can apply tailored sampling patterns to selectively extract photons that yield the most information for particle localization. We demonstrate that off-center sampling at locations predicted by Fisher information utilizes photons most efficiently. When performing localization in a single dimension, optimized off-center sampling patterns gave doubled precision compared to uniform sampling. A ~20% increase in precision compared to uniform sampling can be achieved when a similar off-center pattern is used in 3D localization. This work is the first to thoroughly investigate the photon efficiency of different emission patterns in a diffraction-limited system and achieve higher precision than uniform sampling. The ability to maximize information from the limited number of photons demonstrated here is critical for particle tracking applications in biological samples, where photons may be limited.

Entropy ◽  
2021 ◽  
Vol 23 (5) ◽  
pp. 498
Author(s):  
Chen Zhang ◽  
Kevin Welsher

In this work, we present a 3D single-particle tracking system that can apply tailored sampling patterns to selectively extract photons that yield the most information for particle localization. We demonstrate that off-center sampling at locations predicted by Fisher information utilizes photons most efficiently. When performing localization in a single dimension, optimized off-center sampling patterns gave doubled precision compared to uniform sampling. A ~20% increase in precision compared to uniform sampling can be achieved when a similar off-center pattern is used in 3D localization. Here, we systematically investigated the photon efficiency of different emission patterns in a diffraction-limited system and achieved higher precision than uniform sampling. The ability to maximize information from the limited number of photons demonstrated here is critical for particle tracking applications in biological samples, where photons may be limited.


2010 ◽  
Author(s):  
Ai-Tang Chang ◽  
Yi-Ren Chang ◽  
Sien Chi ◽  
Long Hsu

Author(s):  
Donghee Lee ◽  
Jeonghoon Lee ◽  
Jung Kyung Kim

Fluorescence recovery after photobleaching (FRAP) and single particle tracking (SPT) techniques determine the diffusion coefficient from average diffusive motion of high-concentration molecules and from trajectories of low-concentration single molecules, respectively. Lateral diffusion coefficients measured by FRAP and SPT techniques for the same biomolecule on cell membrane have exhibited inconsistent values across laboratories and platforms with larger diffusion coefficient determined by FRAP, but the sources of the inconsistency have not been investigated thoroughly. Here, we designed an image-based FRAP-SPT system and made a direct comparison between FRAP and SPT for diffusion coefficient of submicron particles with known theoretical values derived from Stokes–Einstein equation in aqueous solution. The combined [Formula: see text]FRAP-SPT technique allowed us to measure the diffusion coefficient of the same fluorescent particle by utilizing both techniques in a single platform and to scrutinize inherent errors and artifacts of FRAP. Our results reveal that diffusion coefficient overestimated by FRAP is caused by inaccurate estimation of the bleaching spot size and can be corrected by simple image analysis. Our [Formula: see text]FRAP-SPT technique can be potentially used for not only cellular membrane dynamics but also for quantitative analysis of the spatiotemporal distribution of the solutes in small scale analytical devices.


Soft Matter ◽  
2021 ◽  
Author(s):  
Katie A. Rose ◽  
Daeyeon Lee ◽  
Russell J. Composto

The effect of static silica particles on the dynamics of quantum dot (QD) nanoparticles grafted with a poly(ethylene glycol) (PEG) brush in hydrogel nanocomposites is investigated using single particle tracking (SPT).


2013 ◽  
Vol 102 (17) ◽  
pp. 173702 ◽  
Author(s):  
Manuel F. Juette ◽  
Felix E. Rivera-Molina ◽  
Derek K. Toomre ◽  
Joerg Bewersdorf

Sign in / Sign up

Export Citation Format

Share Document