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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editor: Gregory W. Faris
  • Vol. 4, Iss. 12 — Nov. 10, 2009

Polarization sensitive, three-dimensional, single-molecule imaging of cells with a double-helix system

Sri Rama Prasanna Pavani, Jennifer G. DeLuca, and Rafael Piestun  »View Author Affiliations


Optics Express, Vol. 17, Issue 22, pp. 19644-19655 (2009)
http://dx.doi.org/10.1364/OE.17.019644


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Abstract

Double-helix point spread function photoactivation-localization microscopy allows three-dimensional (3D) superresolution imaging of objects smaller than the optical diffraction-limit. We demonstrate polarization sensitive detection with 3D super-localization of single-molecules and unveil 3D polarization specific characteristics of single-molecules within the intracellular structure of PtK1 cells expressing photoactivatable green fluorescent protein. The system modulates orthogonal polarization components of single-molecule emissions with a single spatial light modulator and detects them separately with a single detector. Information obtained from the two polarization channels demonstrates polarization based contrast in 3D superresolution imaging. Further, we show that the 3D information from the two channels can be optimally combined to yield up to 30% improvement in localization precision relative to a single polarization channel system.

© 2009 OSA

OCIS Codes
(100.6640) Image processing : Superresolution
(110.4850) Imaging systems : Optical transfer functions
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(260.5430) Physical optics : Polarization
(110.1758) Imaging systems : Computational imaging

ToC Category:
Microscopy

History
Original Manuscript: August 7, 2009
Revised Manuscript: October 5, 2009
Manuscript Accepted: October 7, 2009
Published: October 15, 2009

Virtual Issues
Vol. 4, Iss. 12 Virtual Journal for Biomedical Optics

Citation
Sri Rama Prasanna Pavani, Jennifer G. DeLuca, and Rafael Piestun, "Polarization sensitive, three-dimensional, single-molecule imaging of cells with a 
double-helix system," Opt. Express 17, 19644-19655 (2009)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-17-22-19644


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