Journal
JOURNAL OF MICROSCOPY-OXFORD
Volume 204, Issue -, Pages 99-107Publisher
BLACKWELL SCIENCE LTD
DOI: 10.1046/j.1365-2818.2001.00949.x
Keywords
confocal microscopy; conjugate gradients; deconvolution; fluorescence; Hessian matrix; image restoration; line-search; point spread function; three-dimensional microscopy; wide-field microscopy
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For deconvolution applications in three-dimensional microscopy we derived and implemented a generic, accelerated maximum likelihood image restoration algorithm. A conjugate gradient iteration scheme was used considering either Gaussian or Poisson noise models. Poisson models are better suited to low intensity fluorescent image data: typically, they show smaller restoration errors and smoother results. For the regularization, we modified the standard Tikhonov method. However, the generic design of the algorithm allows for more regularization approaches. The Hessian matrix of the restoration functional was used to determine the step size. We compared restoration error and convergence behaviour between the classical line-search and the Hessian matrix method. Under typical working conditions, the restoration error did not increase over that of the line-search and the speed of convergence did not significantly decrease allowing for a twofold increase in processing speed. To determine the regularization parameter, we modified the generalized cross-validation method. Tests that were done on both simulated and experimental fluorescence wide-field data show reliable results.
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