Abstract

Interferometric detection of the fluorescence emission from a single molecule [interferometric photoactivated localization microscopy (iPALM)] enables a localization accuracy of nanometers in axial localization for 3D superresolution imaging. However, iPALM uses two high-numerical-aperture (NA) objectives in juxtaposition for fluorescence collection (a 4Pi microscope geometry), increasing expense and limiting samples that can be studied. Here, we propose an interferometric single molecule localization microscopy method using a single high-NA objective. The axial position of single molecules can be unambiguously determined from the phase-shifted interference signals with nanometer precision and over a range of $2\lambda$. The use of only one objective simplifies the system configuration and sample mounting. In addition, due to the use of wavefront-splitting interference in our approach, the two parts of the wavefront that eventually merge and interfere with each other travel along nearly equivalent optical paths, which should minimize the effect of drift for long-term 3D superresolution imaging.

© 2014 Optical Society of America

3D superresolution microscopy by supercritical angle detection

Joran Deschamps, Markus Mund, and Jonas Ries
Opt. Express 22(23) 29081-29091 (2014)

4Pi fluorescence detection and 3D particle localization with a single objective

J. Schnitzbauer, R. McGorty, and B. Huang
Opt. Express 21(17) 19701-19708 (2013)

Design and simulation of a snapshot multi-focal interferometric microscope

Kuan He, Xiang Huang, Xiaolei Wang, Seunghwan Yoo, Pablo Ruiz, Itay Gdor, Nicola J. Ferrier, Norbert Scherer, Mark Hereld, Aggelos K. Katsaggelos, and Oliver Cossairt
Opt. Express 26(21) 27381-27402 (2018)

Polarization sensitive, three-dimensional, single-molecule imaging of cells with a double-helix system

Sri Rama Prasanna Pavani, Jennifer G. DeLuca, and Rafael Piestun
Opt. Express 17(22) 19644-19655 (2009)

Three-dimensional biplane spectroscopic single-molecule localization microscopy

Ki-Hee Song, Yang Zhang, Gaoxiang Wang, Cheng Sun, and Hao F. Zhang
Optica 6(6) 709-715 (2019)