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Interferometric three-dimensional single molecule localization microscopy using a single high-numerical-aperture objective.

Authors
  • Zhang, P
  • Goodwin, P M
  • Werner, J H
Type
Published Article
Journal
Applied Optics
Publisher
The Optical Society
Publication Date
Nov 01, 2014
Volume
53
Issue
31
Pages
7415–7421
Identifiers
DOI: 10.1364/AO.53.007415
PMID: 25402907
Source
Medline
License
Unknown

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 2λ. 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.

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