title: Enhanced image reconstruction of three-dimensional fluorescent assays by subtractive structured-light illumination microscopy.
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.
authors.affiliation: School of Electrical and Electronic Engineering, Yonsei University, Seoul, South Korea.
authors.forename: Jong-ryul
authors.initials: JR
authors.lastname: Choi
authors.forename: Donghyun
authors.initials: D
authors.lastname: Kim
mesh_terms: D000464:Alginates; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D008863:Microspheres; D013050:Spectrometry, Fluorescence; D013382:Subtraction Technique
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid