Delpiano J, Jara J, Scheer J, Ramírez O, Ruiz-del-Solar J and S Härtel
Machine Vision and Applications, Volume 23, Issue 4, pp: 675-89

Optical flow (OF) approaches calculate vector fields which determine the apparent velocities of objects in time-varying image sequences. They have been analyzed extensively in computer science using both natural and synthetic video sequences. In life sciences, there is an increasing need to extract kinetic information from temporal image sequences which reveals the interplay between form and function of microscopic biological structures. In this work, we test different variational OF techniques to quantify the displacements of biological objects in 2D fluorescent image sequences. The accuracy of the vector fields is tested for defined displacements of fluorescent point sources in synthetic image series which mimic protein traffic in neuronal dendrites, and for GABABR1 receptor subunits in dendrites of hippocampal neurons. Our results reveal that OF fields predict the movement of fluorescent point sources within an error of 3% for a maximum displacement of 160 nm. Displacement of agglomerated GABABR1 receptor subunits can be predicted correctly for maximum displacements of 640 nm. We further introduce a criteria to derive the optimum parameter combinations for the calculation of the OF fields in experimental images. From these results, temporal sampling frequencies for image acquisition can be derived to guarantee correct motion estimation for biological objects.

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