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J Opt Soc Am A Opt Image Sci Vis. 2012 Oct 1;29(10):2165-73. doi: 10.1364/JOSAA.29.002165.

Enhanced image reconstruction of three-dimensional fluorescent assays by subtractive structured-light illumination microscopy.

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1
School of Electrical and Electronic Engineering, Yonsei University, Seoul, South Korea.

Abstract

We investigate improved image reconstruction of structured light illumination for high-resolution imaging of three-dimensional (3D) cell-based assays. For proof of concept, an in situ fluorescence optical detection system was built with a digital micromirror device as a spatial light modulator, for which phase and tilting angle in a grid pattern were varied to implement specific image reconstruction schemes. Subtractive reconstruction algorithms based on structured light illumination were used to acquire images of fluorescent microbeads deposited as a two-dimensional monolayer or in 3D alginate matrix. We have confirmed that an optical subtraction algorithm improves axial and lateral resolution by effectively removing out-of-focus fluorescence. The results suggest that subtractive image reconstruction can be useful for structured illumination microscopy of broad types of cell-based assays with high image resolution.

PMID:
23201665
[Indexed for MEDLINE]
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