3D microscope imaging robust to restoration artifacts introduced by optically thick specimens

Nurmohammed Patwary; Sharon V. King; Chrysanthe Preza

doi:10.1117/12.2077845

9 March 2015 3D microscope imaging robust to restoration artifacts introduced by optically thick specimens

Nurmohammed Patwary, Sharon V. King, Chrysanthe Preza

Proceedings Volume 9330, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII; 93300O (2015) https://doi.org/10.1117/12.2077845
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

We demonstrate 3D microscope imaging using computational optical sectioning microscopy (COSM) with an engineered point-spread function (PSF) robust to depth-induced spherical aberration (SA). Earlier we demonstrated that wavefront encoding (WFE) using a squared cubic (SQUBIC) phase mask reduces the PSF depth-variance in the presence of SA and that space-invariant (SI) restoration of simulated images using a single WFE-PSF does not lead to artifacts as in the conventional case. In this study, we show experimental verification of our WFE COSM approach. The WFE system used is a commercial microscope with a modified side port imaging path, where a spatial light modulator projects the SQUBIC phase mask on the back focal plane of the imaging lens. High resolution images of a test sample with 6 μm in diameter microspheres embedded in UV-cured optical cement (RI = 1.47) were captured using both the engineered and the conventional imaging paths of the system. The acquired images were restored using a regularized SI expectation maximization algorithm based on Tikhonov-Miller regularization with a roughness penalty. A comparative study quantified in terms of the correlation coefficients between the XZ medial sections of the restored images, from experimental data, shows an 11% reduction in depth sensitivity in the SQUBIC system compared to the conventional system.

Citation Download Citation

Nurmohammed Patwary, Sharon V. King, and Chrysanthe Preza "3D microscope imaging robust to restoration artifacts introduced by optically thick specimens", Proc. SPIE 9330, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII, 93300O (9 March 2015); https://doi.org/10.1117/12.2077845

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