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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
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Tracking Single Proteins in Lipid Bilayers Using Fluorescence Microscopy
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Tracking multiple particles in fluorescence time-lapse microscopy images via probabilistic data association.

William J Godinez, Karl Rohr

    IEEE Transactions on Medical Imaging
    |September 25, 2014
    PubMed
    Summary

    This study introduces a new method for tracking multiple fluorescent particles in microscopy images using probabilistic data association. This approach enhances the analysis of dynamical processes in subcellular and viral structures.

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    Area of Science:

    • Microscopy
    • Biophysics
    • Image Analysis

    Background:

    • Quantitative analysis of dynamical processes relies on tracking subcellular and viral structures in fluorescence microscopy.
    • Accurate tracking of multiple particles, especially those in close proximity, is challenging.

    Purpose of the Study:

    • To develop a robust approach for tracking multiple fluorescent particles in 2-D and 3-D microscopy images.
    • To improve the accuracy and reliability of particle tracking for analyzing dynamical processes.

    Main Methods:

    • Probabilistic data association for tracking multiple fluorescent particles.
    • Combined localization scheme using bottom-up (spot-enhancing filter) and top-down (ellipsoidal sampling) strategies.
    • Integration of Kalman filter with image likelihood for weighted measurements and interacting multiple model algorithm for motion models.

    Main Results:

    • Successfully applied the developed approach to synthetic and real 2-D and 3-D microscopy images.
    • Quantified the performance of the tracking method.
    • Achieved successful application to data from the IEEE ISBI 2012 Particle Tracking Challenge.

    Conclusions:

    • The developed probabilistic data association approach provides accurate and reliable tracking of multiple fluorescent particles.
    • This method enhances the quantitative analysis of dynamical processes in biological imaging.
    • The approach is effective for both synthetic and real microscopy data, including challenging scenarios with closely located objects.