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2 Janelia Publications

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    Looger Lab
    02/01/09 | A bright and photostable photoconvertible fluorescent protein.
    McKinney SA, Murphy CS, Hazelwood KL, Davidson MW, Looger LL
    Nature Methods. 2009 Feb;6(2):131-3. doi: 10.1038/nmeth.1296

    Photoconvertible fluorescent proteins are potential tools for investigating dynamic processes in living cells and for emerging super-resolution microscopy techniques. Unfortunately, most probes in this class are hampered by oligomerization, small photon budgets or poor photostability. Here we report an EosFP variant that functions well in a broad range of protein fusions for dynamic investigations, exhibits high photostability and preserves the approximately 10-nm localization precision of its parent.

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    02/01/09 | Automatic tracking of Escherichia coli in phase-contrast microscopy video.
    Xie J, Khan S, Shah M
    IEEE Transactions on Bio-Medical Engineering. 2009 Feb;56(2):390-9. doi: 10.1109/TBME.2008.2005956

    In this paper, we present an automatic method for estimating the trajectories of Escherichia coli bacteria from in vivo phase-contrast microscopy. To address the low-contrast boundaries in cellular images, an adaptive kernel-based technique is applied to detect cells in each frame. In addition to intensity features, region homogeneity measure and class uncertainty are also applied in this detection technique. To track cells with complex motion, a novel matching gain measure is introduced to cope with the challenges, particularly the presence of low-contrast boundary, the variations of appearance, and the frequent overlapping and occlusion. For multicell tracking over time, an optimal matching strategy is introduced to improve the handling of cell collision and broken trajectories. The results of successful tracking of Escherichia coli from various phase-contrast sequences are reported and compared with manually determined trajectories, as well as those obtained from existing tracking schemes. The stability of the algorithm with different parameter values is also analyzed and discussed.

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