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Proceedings Paper

New results on the single molecule localization problem in two and three dimensions
Author(s): Amir Tahmasbi; E. Sally Ward; Raimund J. Ober
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Paper Abstract

Fluorescence microscopy is an optical microscopy technique which has been extensively used to study specifically- labeled subcellular objects, such as proteins, and their functions. The best possible accuracy with which an object of interest can be localized when imaged using a fluorescence microscope is typically calculated using the Cramer- Rao lower bound (CRLB). The calculation of the CRLB, however, so far relied on an analytical expression for the image of the object. This can pose challenges in practice since it is often difficult to find appropriate analytical models for the images of general objects. Even if an appropriate analytical model is available, the lack of knowledge about the precise values of imaging parameters might also impose difficulties in the calculation of the CRLB. To address these challenges, we have developed an approach that directly uses an experimentally collected image set to calculate the best possible localization accuracy for a general subcellular object in two and three dimensions. In this approach, we fit smoothly connected piecewise polynomials, known as splines, to the experimentally collected image set to provide a continuous model of the object. This continuous model can then be used for the calculation of the best possible localization accuracy.

Paper Details

Date Published: 26 August 2015
PDF: 8 pages
Proc. SPIE 9554, Nanoimaging and Nanospectroscopy III, 955402 (26 August 2015); doi: 10.1117/12.2192008
Show Author Affiliations
Amir Tahmasbi, Texas A&M Univ. (United States)
Texas A&M Health Science Ctr. (United States)
E. Sally Ward, Texas A&M Health Science Ctr. (United States)
Raimund J. Ober, Texas A&M Univ. (United States)
Texas A&M Health Science Ctr. (United States)

Published in SPIE Proceedings Vol. 9554:
Nanoimaging and Nanospectroscopy III
Prabhat Verma; Alexander Egner, Editor(s)

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