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Optics Letters

Optics Letters


  • Editor: Alan E. Willner
  • Vol. 38, Iss. 9 — May. 1, 2013
  • pp: 1521–1523

Single-molecule orientation measurements with a quadrated pupil

Adam S. Backer, Mikael P. Backlund, Matthew D. Lew, and W. E. Moerner  »View Author Affiliations

Optics Letters, Vol. 38, Issue 9, pp. 1521-1523 (2013)

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This Letter presents a means of measuring the dipole orientation of a fluorescent, orientationally fixed single molecule, which uses a specially designed phase mask, termed a “quadrated pupil,” conjugate to the back focal plane of a conventional wide-field microscope. The method leverages the spatial anisotropy of the far-field emission pattern of a dipole emitter and makes this anisotropy amenable to quantitative analysis at the image plane. In comparison to older image-fitting techniques that infer orientation by matching simulations to defocused or excessively magnified images, the quadrated pupil approach is more robust to minor modeling discrepancies and optical aberrations. Precision of 1°–5° is achieved in proof-of-concept experiments for both azimuthal (φ) and polar (θ) angles without defocusing. Since the phase mask is implemented on a liquid-crystal spatial light modulator that may be deactivated without any mechanical perturbation of the sample or imaging system, the technique may be readily integrated into clear aperture imaging studies.

© 2013 Optical Society of America

OCIS Codes
(070.6110) Fourier optics and signal processing : Spatial filtering
(110.4850) Imaging systems : Optical transfer functions
(180.2520) Microscopy : Fluorescence microscopy
(110.1758) Imaging systems : Computational imaging

ToC Category:
Imaging Systems

Original Manuscript: January 7, 2013
Revised Manuscript: March 11, 2013
Manuscript Accepted: March 13, 2013
Published: April 29, 2013

Virtual Issues
Vol. 8, Iss. 6 Virtual Journal for Biomedical Optics

Adam S. Backer, Mikael P. Backlund, Matthew D. Lew, and W. E. Moerner, "Single-molecule orientation measurements with a quadrated pupil," Opt. Lett. 38, 1521-1523 (2013)

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