Title :

Design of experiments for the optimization of SOFI super-resolution microscopy imaging

Author(s) :

Cevoli, Dario [Auteur]
Catholic University of Leuven = Katholieke Universiteit Leuven [KU Leuven]
Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement - UMR 8516 [LASIRE]
Vitale, Raffaele [Auteur]
Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement (LASIRE) - UMR 8516
Vandenberg, W. [Auteur]
Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement - UMR 8516 [LASIRE]
Hugelier, S. [Auteur]
Department of Pharmaceutical and Pharmacological Sciences, KU Leuven
Van Den Eynde, R. [Auteur]
Catholic University of Leuven = Katholieke Universiteit Leuven [KU Leuven]
Dedecker, P. [Auteur]
Catholic University of Leuven = Katholieke Universiteit Leuven [KU Leuven]
Ruckebusch, Cyril [Auteur]
Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement (LASIRE) - UMR 8516
Vandenberg, W. [Auteur]

Journal title :

Biomed. Opt. Express

Abbreviated title :

Biomed. Opt. Express

Volume number :

12

Pages :

2617-2630

Publication date :

2021-06-10

ISSN :

2156-7085

HAL domain(s) :

Chimie/Chimie théorique et/ou physique

English abstract : [en]

Super-resolution optical fluctuation imaging (SOFI) is a well-known super-resolution technique appreciated for its versatility and broad applicability. However, even though an extended theoretical description is available, ...
Show more >Super-resolution optical fluctuation imaging (SOFI) is a well-known super-resolution technique appreciated for its versatility and broad applicability. However, even though an extended theoretical description is available, it is still not fully understood how the interplay between different experimental parameters influences the quality of a SOFI image. We investigated the relationship between five experimental parameters (measurement time, on-time ton, off-time toff, probe brightness, and out of focus background) and the quality of the super-resolved images they yielded, expressed as Signal to Noise Ratio (SNR). Empirical relationships were modeled for second- and third-order SOFI using data simulated according to a D-Optimal design of experiments, which is an ad-hoc design built to reduce the experimental load when the total number of trials to be conducted becomes too high for practical applications. This approach proves to be more reliable and efficient for parameter optimization compared to the more classical parameter by parameter approach. Our results indicate that the best image quality is achieved for the fastest emitter blinking (lowest ton and toff), lowest background level, and the highest measurement duration, while the brightness variation does not affect the quality in a statistically significant way within the investigated range. However, when the ranges spanned by the parameters are constrained, a different set of optimal conditions may arise. For example, for second-order SOFI, we identified situations in which the increase of toff can be beneficial to SNR, such as when the measurement duration is long enough. In general, optimal values of ton and toff have been found to be highly dependent from each other and from the measurement duration.Show less >

Language :

Anglais

Audience :

Internationale

Popular science :

Non

Administrative institution(s) :

Université de Lille
CNRS

Collections :

• Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement (LASIRE) - UMR 8516

Submission date :

2024-02-28T22:40:00Z
2024-03-15T12:39:16Z