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Polo-like kinases (Plks) perform crucial functions in cell-cycle progression and multiple stages of mitosis. Plks are characterized by a C-terminal noncatalytic region containing two tandem Polo boxes, termed the Polo-box domain (PBD), which has recently been implicated in phosphodependent substrate targeting. We show that the PBDs of human, Xenopus, and(More)
Estimating the location of single molecules from microscopy images is a key step in many quantitative single molecule data analysis techniques. Different algorithms have been advocated for the fitting of single molecule data, particularly the nonlinear least squares and maximum likelihood estimators. Comparisons were carried out to assess the performance of(More)
Super-localization microscopy encompasses techniques that depend on the accurate localization of individual molecules from generally low-light images. The obtainable localization accuracies, however, are ultimately limited by the image detector's pixelation and noise. We present the ultrahigh accuracy imaging modality (UAIM), which allows users to obtain(More)
A three-dimensional (3D) resolution measure for the conventional optical microscope is introduced which overcomes the drawbacks of the classical 3D (axial) resolution limit. Formulated within the context of a parameter estimation problem and based on the Cramer-Rao lower bound, this 3D resolution measure indicates the accuracy with which a given distance(More)
From an acquired image, single molecule microscopy makes possible the determination of the distance separating two closely spaced biomolecules in three-dimensional (3D) space. Such distance information can be an important indicator of the nature of the biomolecular interaction. Distance determination, however, is especially difficult when, for example, the(More)
Different techniques have been advocated for estimating single molecule locations from microscopy images. The question arises as to which technique produces the most accurate results. Various factors, e.g. the stochastic nature of the photon emission/detection process, extraneous additive noise, pixelation, etc., result in the estimated single molecule(More)
Technological advances in both hardware and software have made possible the realization of sophisticated biological imaging experiments using the optical microscope. As a result, modern microscopy experiments are capable of producing complex image datasets. For a given data analysis task, the images in a set are arranged, based on the requirements of the(More)
An information-theoretic three-dimensional (3D) resolution measure for the optical microscope is introduced. Based on the Cramer-Rao inequality, this resolution measure specifies a lower bound on the accuracy with which a given distance separating two objects in 3D space can be estimated from the acquired image. Useful in many applications, accurate(More)
Methods: Twenty-four Sprague Dawley rats were divided into 2 groups. All underwent left sciatic nerve injury, leaving a 10mm gap. The injury was repaired using either: 1) Reverse autograft – the 10mm cut segment was oriented 180° and used to coapt the proximal and distal ends or 2) UBM conduit-the 10mm nerve gap was bridged with UBM conduit. Validated(More)
Using a point spread function (PSF) to localize a point-like object, such as a fluorescent molecule or microsphere, represents a common task in single molecule microscopy image data analysis. The localization may differ in purpose depending on the application or experiment, but a unifying theme is the importance of being able to closely recover the true(More)