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Proceedings Paper

Single-molecule orientation measurements with a quadrated pupil
Author(s): Adam S. Backer; Mikael P. Backlund; Matthew D. Lew; Alexander R. Diezmann; Steffen J. Sahl; W. E. Moerner
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Paper Abstract

We present a means of measuring the dipole orientation of a fluorescent, rotationally fixed single molecule (SM), using a specially designed phase mask, termed a “quadrated pupil,” conjugate to the back focal plane of a conventional widefield microscope. In comparison to 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, defocus, and optical aberrations. Precision on the order of 1°-5° is achieved in proofof- concept experiments for both azimuthal (φ) and polar (θ) angles. Since the phase mask is implemented on a liquid-crystal spatial light modulator (SLM) that may be deactivated without any mechanical perturbation of the sample or imaging system, the technique may be readily integrated into conventional imaging studies.

Paper Details

Date Published: 4 March 2014
PDF: 6 pages
Proc. SPIE 8950, Single Molecule Spectroscopy and Superresolution Imaging VII, 89500L (4 March 2014); doi: 10.1117/12.2042097
Show Author Affiliations
Adam S. Backer, Stanford Univ. (United States)
Mikael P. Backlund, Stanford Univ. (United States)
Matthew D. Lew, Stanford Univ. (United States)
Alexander R. Diezmann, Stanford Univ. (United States)
Steffen J. Sahl, Stanford Univ. (United States)
W. E. Moerner, Stanford Univ. (United States)


Published in SPIE Proceedings Vol. 8950:
Single Molecule Spectroscopy and Superresolution Imaging VII
Jörg Enderlein; Ingo Gregor; Zygmunt Karol Gryczynski; Rainer Erdmann; Felix Koberling, Editor(s)

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