Expand this Topic clickable element to expand a topic
Skip to content
Optica Publishing Group

Single-molecule orientation measurements with a quadrated pupil

Not Accessible

Your library or personal account may give you access

Abstract

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

Full Article  |  PDF Article
More Like This
Three dimensional single molecule localization using a phase retrieved pupil function

Sheng Liu, Emil B. Kromann, Wesley D. Krueger, Joerg Bewersdorf, and Keith A. Lidke
Opt. Express 21(24) 29462-29487 (2013)

Determination of the three-dimensional orientation of single molecules

Matthew R. Foreman, Carlos Macías Romero, and Peter Török
Opt. Lett. 33(9) 1020-1022 (2008)

References

You do not have subscription access to this journal. Citation lists with outbound citation links are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Cited By

You do not have subscription access to this journal. Cited by links are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Figures (2)

You do not have subscription access to this journal. Figure files are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Tables (1)

You do not have subscription access to this journal. Article tables are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Equations (4)

You do not have subscription access to this journal. Equations are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription

Metrics

Select as filters


Select Topics Cancel
© Copyright 2022 | Optica Publishing Group. All Rights Reserved