Single-molecule orientation measurements with a quadrated pupil

Adam S. Backer, Mikael P. Backlund, Matthew D. Lew, W. E. Moerner

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review


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 imagefitting 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 on the order of 1o-5o is achieved in proof-of-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.

Original languageEnglish
Title of host publicationImaging Systems and Applications, ISA 2013
PublisherOptical Society of America (OSA)
ISBN (Print)9781557529756
StatePublished - 2013
EventImaging Systems and Applications, ISA 2013 - Arlington, VA, United States
Duration: Jun 23 2013Jun 27 2013

Publication series

NameOptics InfoBase Conference Papers
ISSN (Electronic)2162-2701


ConferenceImaging Systems and Applications, ISA 2013
Country/TerritoryUnited States
CityArlington, VA


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