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Precise 3-D localization of fluorescent probes without numerical Ting Sun and Sean B. Andersson
 

Summary: Precise 3-D localization of fluorescent probes without numerical
fitting
Ting Sun and Sean B. Andersson
Abstract--We present an analytical algorithm for determining
the position of a fluorescent probe in three dimensions (3-D) from
a collection of measurements taken at different locations. This
algorithm, fluoroBancroft, relies on the fact that the intensity
point spread function depends only on the radial distance between
the center of the focal point and the position of the fluorescent
probe. We present a simulation study comparing the performance
of the algorithm to the standard technique of fitting the data to
a Gaussian profile. Our results indicate that in the 3-D case, the
fluoroBancroft algorithm is able to localize the probe with an
accuracy on the order of tens of nanometers using less than
ten measurements (pixels). The Gaussian fitting procedure is
unable to locate the probe even when using 36 measurements
(the maximum number of measurements used in the simulations).
Moreover the new technique is typically two orders of magnitude
faster than the Gaussian fitting approach in terms of computation
time. These results indicate the fluoroBancroft algorithm can be

  

Source: Andersson, Sean B. - Department of Aerospace and Mechanical Engineering, Boston University

 

Collections: Engineering