Application of image restoration methods for confocal fluorescence microscopy

Geert M. P. van Kempen; Lucas J. van Vliet; Peter J. Verveer

doi:10.1117/12.271257

10 April 1997 Application of image restoration methods for confocal fluorescence microscopy

Geert M. P. van Kempen, Lucas J. van Vliet, Peter J. Verveer

Author Affiliations +

Proceedings Volume 2984, Three-Dimensional Microscopy: Image Acquisition and Processing IV; (1997) https://doi.org/10.1117/12.271257
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States

Abstract

The analysis of the three-dimensional structure of tissue, cells and cellular constituents play a major role in biomedical research. Three-dimensional images, acquired by confocal fluorescence microscopes play a key role in this analysis. However, the imaging properties of these microscopes give rise to diffraction-induced blurring phenomena. These distortions hamper subsequent quantitative analysis. Therefore, restoration algorithms that invert these distortions will improve these analyses. We have tested the performances of the Richardson-Lucy, and the ICTM algorithm in a simulation experiment and found a strong dependency of their performances on the signal-to-noise ratio of the image. We propose a pre-filtering to reduce the noise in the image without hampering the object. We have applied a Gaussian filter and a median filter prior to the restoration, and compensate for this extra blurring of the Gaussian in the restoration procedure. We show how this pre- filtering improves the performance of the restoration algorithms. The experiments were performed on spheres convolved with a confocal point spread function and distorted with Poisson noise.

Citation Download Citation

Geert M. P. van Kempen, Lucas J. van Vliet, and Peter J. Verveer "Application of image restoration methods for confocal fluorescence microscopy", Proc. SPIE 2984, Three-Dimensional Microscopy: Image Acquisition and Processing IV, (10 April 1997); https://doi.org/10.1117/12.271257

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