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J Microsc. 2005 Jan;217(Pt 1):93-108.

Noise suppression of point spread functions and its influence on deconvolution of three-dimensional fluorescence microscopy image sets.

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1
Center for Immunology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.

Abstract

The point spread function (PSF) is of central importance in the image restoration of three-dimensional image sets acquired by an epifluorescent microscope. Even though it is well known that an experimental PSF is typically more accurate than a theoretical one, the noise content of the experimental PSF is often an obstacle to its use in deconvolution algorithms. In this paper we apply a recently introduced noise suppression method to achieve an effective noise reduction in experimental PSFs. We show with both simulated and experimental three-dimensional image sets that a PSF that is smoothed with this method leads to a significant improvement in the performance of deconvolution algorithms, such as the regularized least-squares algorithm and the accelerated Richardson-Lucy algorithm.

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