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Performance of optical flow techniques for motion analysis of fluorescent point signals in confocal microscopy

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Abstract

Optical flow 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 optical flow 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 optical flow 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. Based on these results, we introduce a criteria to derive the optimum parameter combinations for the calculation of the optical flow 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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Authors and Affiliations

  1. Department of Electrical Engineering, Faculty of Physical and Mathematical Sciences, University of Chile, Av. Beauchef 850, Santiago, Chile

    José Delpiano & Javier Ruiz-del-Solar

  2. Faculty of Engineering and Applied Sciences, University of the Andes, Av. San Carlos de Apoquindo 2200, Santiago, Chile

    José Delpiano

  3. Laboratory for Scientific Image Analysis (SCIAN-Lab) at the Program of Anatomy and Developmental Biology and the Biomedical Neuroscience Institute BNI, ICBM, Faculty of Medicine, University of Chile, Independencia 1027, Santiago, Chile

    Jorge Jara, Jan Scheer, Omar A. Ramírez & Steffen Härtel

  4. Department of Computer Sciences, Faculty of Physical and Mathematical Sciences, University of Chile, Av. Beauchef 850, Santiago, Chile

    Jorge Jara

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  1. José Delpiano
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  2. Jorge Jara
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  5. Javier Ruiz-del-Solar
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Correspondence to Steffen Härtel.

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Delpiano, J., Jara, J., Scheer, J. et al. Performance of optical flow techniques for motion analysis of fluorescent point signals in confocal microscopy. Machine Vision and Applications 23, 675–689 (2012). https://doi.org/10.1007/s00138-011-0362-8

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  • DOI: https://doi.org/10.1007/s00138-011-0362-8

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