Visualizing stable features in live cell nucleus for evaluation of the cell global motion compensation

The compensation of cell motion is an important step in single-particle tracking analysis of live cells. This step is required in most of the cases, since the movement of subcellular foci is superimposed by the movement and deformation of the cell, while only the local motion of the foci is important to be analysed. The cell motion and deformation compensation is usually performed by means of image registration. There are a number of approaches with different models and properties presented in the literature that perform cell image registration. However, the evaluation of the registration approach quality on real data is a tricky problem due to the fact that some stable features in the images with a priori no local motion are needed. In this paper we propose a methodology for creating live cell nuclei image sequences with stable features imposed. The features are defined using the regions of fluorescence bleaching invoked by the UV laser. Data with different deformations are acquired and can be used for evaluation of the cell image registration methods. Along with that, we describe an image analysis technique and a metric that can characterize the quality of the method quantitatively. The proposed methodology allows building a ground truth dataset for testing and thoroughly evaluating cell image registration methods.