Localization beyond the diffraction limit (Conference Presentation)

Petar N. Petrov; Yoav Shechtman; William E. Moerner

doi:10.1117/12.2251307

24 April 2017 Localization beyond the diffraction limit (Conference Presentation)

Petar N. Petrov, Yoav Shechtman, William E. Moerner

Proceedings Volume 10071, Single Molecule Spectroscopy and Superresolution Imaging X; 1007108 (2017) https://doi.org/10.1117/12.2251307
Event: SPIE BiOS, 2017, San Francisco, California, United States

Abstract

Point spread function (PSF) engineering has extended far-field localization microscopy into three dimensions by encoding the axial position of each emitter into the shape of its image on the detector. By fitting the observed PSF to a model function, one can extract position information with sub-diffraction precision. However, in practice this procedure is often complicated by optical aberrations present in the imaging system, which distort the shape of the observed PSF relative to the model function. The mismatch between the model and observed PSFs can limit the accuracy and precision achieved by the localization procedure. Here, we present a simple method to experimentally improve the model PSF by phase retrieval of the pupil function of the imaging system using a set of images of an isolated emitter at different displacements from the focal plane. The pupil function is estimated by adding a phase term consisting of a combination of Zernike modes to the theoretical electric field at the back focal plane of the microscope. The amplitudes of the Zernike modes are determined by maximizing the likelihood function over all pixels in the experimental data set. Importantly, since all data is taken with the phase mask in place, we account for any aberrations it introduces. Using the resulting pupil function, we generate a model PSF which is significantly improved over the theoretical model in both the accuracy and precision of experimental emitter localizations. We also provide a MATLAB package which performs the entire fitting procedure, from phase retrieval to single-emitter localization.

Conference Presentation

Citation Download Citation

Petar N. Petrov, Yoav Shechtman, and William E. Moerner "Localization beyond the diffraction limit (Conference Presentation)", Proc. SPIE 10071, Single Molecule Spectroscopy and Superresolution Imaging X, 1007108 (24 April 2017); https://doi.org/10.1117/12.2251307

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

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: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
PRESENTATION

WATCH
PRESENTATION SAVE TO MY LIBRARY

GET CITATION

KEYWORDS

Point spread functions

Diffraction

Imaging systems

Phase retrieval

3D image processing

3D modeling

Computer programming

Show All Keywords

Keywords/Phrases

Search In:

Publication Years