Single-molecule orientation measurements with a quadrated pupil

Adam S. Backer; Mikael P. Backlund; Matthew D. Lew; Alexander R. Diezmann; Steffen J. Sahl; W. E. Moerner

doi:10.1117/12.2042097

4 March 2014 Single-molecule orientation measurements with a quadrated pupil

Adam S. Backer, Mikael P. Backlund, Matthew D. Lew, Alexander R. Diezmann, Steffen J. Sahl, W. E. Moerner

Author Affiliations +

Proceedings Volume 8950, Single Molecule Spectroscopy and Superresolution Imaging VII; 89500L (2014) https://doi.org/10.1117/12.2042097
Event: SPIE BiOS, 2014, San Francisco, California, United States

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.

Citation Download Citation

Adam S. Backer, Adam S. Backer, Mikael P. Backlund, Mikael P. Backlund, Matthew D. Lew, Matthew D. Lew, Alexander R. Diezmann, Alexander R. Diezmann, Steffen J. Sahl, Steffen J. Sahl, W. E. Moerner, W. E. Moerner, "Single-molecule orientation measurements with a quadrated pupil", Proc. SPIE 8950, Single Molecule Spectroscopy and Superresolution Imaging VII, 89500L (4 March 2014); doi: 10.1117/12.2042097; https://doi.org/10.1117/12.2042097

ACCESS THE FULL ARTICLE

PERSONAL SIGN IN
Full access may be available with your subscription

Email or Username Forgot your username?

Password Forgot your password?

Show

Remember Email/Username on this computer

Remember Password

No SPIE account? Create an account
or Institutional Sign In via Shibboleth

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $14.40

Non-members: $18.00 ADD TO CART

Show All Keywords

Keywords/Phrases

Search In:

Publication Years