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Proceedings Paper

Preprocessing method to correct illumination pattern in sinusoidal-based structured illumination microscopy
Author(s): H. Shabani; A. Doblas; G. Saavedra; C. Preza
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Paper Abstract

The restored images in structured illumination microscopy (SIM) can be affected by residual fringes due to a mismatch between the illumination pattern and the sinusoidal model assumed by the restoration method. When a Fresnel biprism is used to generate a structured pattern, this pattern cannot be described by a pure sinusoidal function since it is distorted by an envelope due to the biprism’s edge. In this contribution, we have investigated the effect of the envelope on the restored SIM images and propose a computational method in order to address it. The proposed approach to reduce the effect of the envelope consists of two parts. First, the envelope of the structured pattern, determined through calibration data, is removed from the raw SIM data via a preprocessing step. In the second step, a notch filter is applied to the images, which are restored using the well-known generalized Wiener filter, to filter any residual undesired fringes. The performance of our approach has been evaluated numerically by simulating the effect of the envelope on synthetic forward images of a 6-μm spherical bead generated using the real pattern and then restored using the SIM approach that is based on an ideal pure sinusoidal function before and after our proposed correction method. The simulation result shows 74% reduction in the contrast of the residual pattern when the proposed method is applied. Experimental results from a pollen grain sample also validate the proposed approach.

Paper Details

Date Published: 23 February 2018
PDF: 6 pages
Proc. SPIE 10499, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV, 104991Z (23 February 2018); doi: 10.1117/12.2291013
Show Author Affiliations
H. Shabani, The Univ. of Memphis (United States)
A. Doblas, The Univ. of Memphis (United States)
G. Saavedra, Univ. de València (Spain)
C. Preza, The Univ. of Memphis (United States)

Published in SPIE Proceedings Vol. 10499:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV
Thomas G. Brown; Carol J. Cogswell; Tony Wilson, Editor(s)

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