On the extended depth of focus algorithms for bright field microscopy

Microscopes offer a limited depth of focus which precludes the observation of a complete image of a three-dimensional (3D) object in a single view. Investigations, by a variety of researchers, have led to the development of extended depth of focus algorithms for serial optical slices of microscopic 3D objects in recent years. However, to date, no quantitative comparison of the different algorithms has been performed, generally leaving the evaluation to the subjective qualitative appreciation of the observer. In this paper we use three different tests for extended depth of focus algorithm evaluation and test 10 different algorithms, some of them have been adapted (by us) for a series of optical slices. However, the main contribution of the paper is a new improved algorithm for computing the extended depth of focus.