Evaluation of image sequence additive decomposition algorithms for membrane analysis in fluorescence video-microscopy

A wide range of proteins viewed using xFP probes in fluorescence video-microscopy shows a cytosolic state diffusing slowly and a membrane state eventually moving as vesicles or small tubules along the cell cytoskeleton network. Due to the property of fluorescence, the measured intensity is the sum of the contributions of these two components. In this study we perform an evaluation of different methods for the automatic decomposition of the two additive components/states. Four algorithms are studied: (i) gray-scale opening (a.k.a. ldquorolling ballrdquo); (ii) wavelet-based detection combined with image interpolation; (iii) Conditional Random Field-based detection; (iv) computational geometry for temporal signal analysis. The evaluation protocol, related to the actual use of image sequence decomposition, includes a qualitative evaluation on real image sequences by experts and a quantitative evaluation on simulated sequences that mimic real images.