Tracking of multiple fluorescent biological objects in three dimensional video microscopy

A method which allows for the first time to perform automatically the detection and the tracking of microscopic objects directly from three dimensional image data is presented. It enables to analyse biological moving objects in three dimensional fluorescence image sequences coming from biological immunomicroscopy experiments, and get quantitative data such as the number of objects, their position, movement phases and speed. After a detection step is performed through the multiscale analysis of images using a shift-invariant wavelet transform, the tracking is achieved using a Kalman filter and an association which enable the position of the moving objects to be predicted, refined and updated. Trajectories are analysed in terms of different parameters relevant for the motility analysis of biological objects.