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

Biomedical Optics Express
Dario CevoliCyril Ruckebusch

Abstract

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, whil...Continue Reading

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Citations

Nov 10, 2021·Analytical Chemistry·Jinkyoung ChungDoory Kim

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