Out of focus background subtraction for fast structured illumination super resolution microscopy of optically thick samples

Abstract : We propose a structured illumination microscopy method to combine super resolution and optical sectioning in three-dimensional (3D) samples that allows the use of two-dimensional (2D) data processing. Indeed, obtaining super-resolution images of thick samples is a difficult task if low spatial frequencies are present in the in-focus section of the sample, as these frequencies have to be distinguished from the out-of-focus background. A rigorous treatment would require a 3D reconstruction of the whole sample using a 3D point spread function and a 3D stack of structured illumination data. The number of raw images required, 15 per optical section in this case, limits the rate at which high-resolution images can be obtained. We show that by a succession of two different treatments of structured illumination data we can estimate the contrast of the illumination pattern and remove the out-of-focus content from the raw images. After this cleaning step, we can obtain super-resolution images of optical sections in thick samples using a two-beam harmonic illumination pattern and a limited number of raw images. This two-step processing makes it possible to obtain super resolved optical sections in thick samples as fast as if the sample was two-dimensional.
Type de document :
Article dans une revue
Journal of Microscopy, Wiley, 2015, 259 (3), pp.257-268 〈10.1111/jmi.12259〉
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https://hal.archives-ouvertes.fr/hal-01225859
Contributeur : François Orieux <>
Soumis le : vendredi 6 novembre 2015 - 20:56:55
Dernière modification le : vendredi 14 décembre 2018 - 01:28:20

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Pierre Vermeulen, H Zhan, François Orieux, Olivo-Marin Jean-Christophe, Zsolt Lenkei, et al.. Out of focus background subtraction for fast structured illumination super resolution microscopy of optically thick samples. Journal of Microscopy, Wiley, 2015, 259 (3), pp.257-268 〈10.1111/jmi.12259〉. 〈hal-01225859〉

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