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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 18, Iss. 7 — Mar. 29, 2010
  • pp: 6563–6576

Precision analysis for standard deviation measurements of immobile single fluorescent molecule images

Michael C. DeSantis, Shawn H. DeCenzo, Je-Luen Li, and Y. M. Wang  »View Author Affiliations

Optics Express, Vol. 18, Issue 7, pp. 6563-6576 (2010)

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Standard deviation measurements of intensity profiles of stationary single fluorescent molecules are useful for studying axial localization, molecular orientation, and a fluorescence imaging system’s spatial resolution. Here we report on the analysis of the precision of standard deviation measurements of intensity profiles of single fluorescent molecules imaged using an EMCCD camera. We have developed an analytical expression for the standard deviation measurement error of a single image which is a function of the total number of detected photons, the background photon noise, and the camera pixel size. The theoretical results agree well with the experimental, simulation, and numerical integration results. Using this expression, we show that single-molecule standard deviation measurements offer nanometer precision for a large range of experimental parameters.

© 2010 Optical Society of America

OCIS Codes
(100.6640) Image processing : Superresolution
(100.6890) Image processing : Three-dimensional image processing
(110.2960) Imaging systems : Image analysis
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Image Processing

Original Manuscript: January 25, 2010
Revised Manuscript: March 1, 2010
Manuscript Accepted: March 2, 2010
Published: March 15, 2010

Virtual Issues
Vol. 5, Iss. 7 Virtual Journal for Biomedical Optics

Michael C. DeSantis, Shawn H. DeCenzo, Je-Luen Li, and Y. M. Wang, "Precision analysis for standard deviation measurements of immobile single fluorescent molecule images," Opt. Express 18, 6563-6576 (2010)

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