title: Experimental validation of a customized phase mask designed to enable efficient computational optical sectioning microscopy through wavefront encoding.
abstract: In this paper, wavefront-encoded (WFE) computational optical sectioning microscopy (COSM) using a fabricated square cubic (SQUBIC) phase mask, designed to render the system less sensitive to depth-induced aberration, is investigated. The WFE-COSM system is characterized by a point spread function (PSF) that does not vary as rapidly with imaging depth compared to the conventional system. Thus, in WFE-COSM, image restoration from large volumes can be achieved using computationally efficient space-invariant (SI) algorithms, thereby avoiding the use of depth-variant algorithms. The fabricated SQUBIC phase mask was first evaluated and found to have a 75% fidelity compared to the theoretical design; it was then integrated in a commercial wide-field microscope to implement a WFE-COSM system. Evaluation of the WFE-COSM system is demonstrated with comparative studies of theoretical and experimental PSFs and simulated and measured images of spherical shells located at different depths in a test sample. These comparisons show that PSF and imaging models capture major trends in experimental data with a 99% correlation between forward image intensity distribution in experimental and simulated images of spherical shells. Our experimental SI restoration results demonstrate that the WFE-COSM system achieves more than a twofold performance improvement over the conventional system of up to a 65 μm depth below the coverslip investigated in this study.
authors.forename: Nurmohammed
authors.initials: N
authors.lastname: Patwary
authors.forename: Hasti
authors.initials: H
authors.lastname: Shabani
authors.forename: Ana
authors.initials: A
authors.lastname: Doblas
authors.forename: Genaro
authors.initials: G
authors.lastname: Saavedra
authors.forename: Chrysanthe
authors.initials: C
authors.lastname: Preza