A design of experiments for the optimization of SOFI super-resolution microscopy imaging

Posted on 07.04.2021 - 15:40
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 and the quality of the super-resolved images they yielded, expressed as SNR. Empirical relationship were modeled for second- and third-order SOFI using data simulated according to a D-Optimal design of experiments. This approach provides to be more reliable and efficient for parameter optimization compared to the more classical parameter by parameter approach. Our results indicate that the best quality is achieved for the fastest emitter blinking (lowest 푡on and 푡off), lowest background level, and the highest measurement duration, while the brightness variation does not affect the image 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 푡off can be beneficial to SNR, such as when the measurement duration is long enough. In general, optimal values of 푡on and 푡off have been found to be highly dependent from each other and from the measurement duration.


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Cevoli, Dario; Vitale, Raffaele; Vandenberg, Wim; hugelier, siewert; Van Den Eynde, Robin; Dedecker, Peter; et al. (2021): A design of experiments for the optimization of SOFI super-resolution microscopy imaging. The Optical Society. Collection. https://doi.org/10.6084/m9.figshare.c.5357408.v2


Biomedical Optics Express


Dario Cevoli
Raffaele Vitale
Wim Vandenberg
siewert hugelier
Robin Van Den Eynde
Peter Dedecker
Cyril Ruckebusch


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