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

Jong Ryul Choi, Donghyun Kim

Research output: Contribution to journalArticle

10 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2165-2173
Number of pages9
JournalJournal of the Optical Society of America A: Optics and Image Science, and Vision
Volume29
Issue number10
DOIs
Publication statusPublished - 2012 Oct 1

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image reconstruction
Image reconstruction
Assays
Microscopic examination
Lighting
illumination
microscopy
Fluorescence
Digital devices
fluorescence
high resolution
Alginate
Optical resolving power
image resolution
light modulators
Image resolution
cells
subtraction
Monolayers
phase shift

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Computer Vision and Pattern Recognition

Cite this

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