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30 March 2020 Can phase masks extend depth-of-field in localization microscopy? (Conference Presentation)
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Abstract
Co-design consists in optimizing an imaging system taking into account the model of image formation, the properties of the optics and the method of information extraction. This methodology is used in biology to observe, with nanometric resolutions, single molecules using fluorescence microscope. This super-resolution imaging technique makes it possible to estimate very accurately the position of an isolated fluorescent particle in the field of the camera beyond the diffraction limit. Today, biologists try to image thicker and less transparent samples. There is then, a need to extend depth-of-field of fluorescence microscope so that single-molecule location accuracy is as invariant as possible by defocalisation. For several years, our team has co-designed phase masks placed in the pupil of a lens to extend its depth-of-field. These masks produce a relatively blurred image, but its quality is independent of the axial position of the object. A unique deconvolution process is then applied to reconstruct the objects at all depths. Optimization of the phase function of the mask is therefore based on an image quality criterion after deconvolution. We propose here to co-design these phase masks for a fluorescence microscope in order to generate a point-spread-function which location accuracy is invariant along the optical axis. From a theoretical point of view, the originality lies in the fact that the optimization criterion is no longer the image quality after deconvolution but the localization accuracy of single-molecule in the plane. Our mask optimization criterion is therefore based on Cramer-Rao lower bound and the associated single-molecule localization post-processing stage is designed such as its accuracy reaches this lower bound. Our results validate this optimization methodology and show that optimal masks can significantly increase the depth of exploration of single-molecule imaging techniques.
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© (2020) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Olivier Lévêque, Caroline Kulcsár, Hervé Sauer, Antony Lee, Pierre Bon, Laurent Cognet, and François Goudail "Can phase masks extend depth-of-field in localization microscopy? (Conference Presentation)", Proc. SPIE 11351, Unconventional Optical Imaging II, 113510E (30 March 2020); https://doi.org/10.1117/12.2558426
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