Open AccessInterferometric three-dimensional single molecule localization microscopy using a single high-numerical-aperture objective
Author(s) -
P. Zhang,
Peter M. Goodwin,
James H. Werner
Publication year - 2014
Publication title -
applied optics
Language(s) - English
Resource type - Journals
ISSN - 0003-6935
DOI - 10.1364/ao.53.007415
Subject(s) - optics , wavefront , interferometry , numerical aperture , microscopy , photoactivated localization microscopy , interference microscopy , microscope , materials science , optical microscope , adaptive optics , physics , super resolution microscopy , wavelength , scanning confocal electron microscopy , scanning electron microscope
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.