High-resolution image reconstruction in fluorescence microscopy with patterned excitation

Appl Opt. 2006 Jul 10;45(20):5037-45. doi: 10.1364/ao.45.005037.

Abstract

We discuss data treatment strategies in structured illumination microscopy, using simulated and experimental data. In the setup, the illumination pattern is generated by projecting a movable pinhole mask into the sample, and a wide-field fluorescence microscope image is acquired for each pattern position. The structured illumination data obtained from a two-dimensional illumination pattern can be treated by projection strategies such as in video confocal microscopy (sum, maximum, maximum minus minimum, and superconfocal), by a scaled subtraction of the out-of-focus estimate, or by a modified version of the Fourier-space treatment, as is known for data from one-dimensional structured illumination. We investigate the influence of some data processing strategies on unwanted effects such as residual patterning and local deviations from linearity in the reconstructed intensity.

Publication types

  • Evaluation Study

MeSH terms

  • Algorithms*
  • Artifacts*
  • Image Enhancement / methods*
  • Image Interpretation, Computer-Assisted / methods*
  • Information Storage and Retrieval / methods
  • Microscopy, Fluorescence / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity