In vivo three-dimensional superresolution fluorescence tracking using a double-helix point spread function

Matthew D. Lew, Michael A. Thompson, Majid Badieirostami, W. E. Moerner

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

19 Scopus citations

Abstract

The point spread function (PSF) of a widefield fluorescence microscope is not suitable for three-dimensional superresolution imaging. We characterize the localization precision of a unique method for 3D superresolution imaging featuring a double-helix point spread function (DH-PSF). The DH-PSF is designed to have two lobes that rotate about their midpoint in any transverse plane as a function of the axial position of the emitter. In effect, the PSF appears as a double helix in three dimensions. By comparing the Cramer-Rao bound of the DH-PSF with the standard PSF as a function of the axial position, we show that the DH-PSF has a higher and more uniform localization precision than the standard PSF throughout a 2 μm depth of field. Comparisons between the DH-PSF and other methods for 3D superresolution are briefly discussed. We also illustrate the applicability of the DH-PSF for imaging weak emitters in biological systems by tracking the movement of quantum dots in glycerol and in live cells.

Original languageEnglish
Title of host publicationSingle Molecule Spectroscopy and Imaging III
DOIs
StatePublished - 2010
EventSingle Molecule Spectroscopy and Imaging III - San Francisco, CA, United States
Duration: Jan 23 2010Jan 25 2010

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume7571
ISSN (Print)1605-7422

Conference

ConferenceSingle Molecule Spectroscopy and Imaging III
Country/TerritoryUnited States
CitySan Francisco, CA
Period01/23/1001/25/10

Keywords

  • Fluorescence microscopy
  • Quantum dot
  • Single-particle tracking
  • Superresolution
  • Three-dimensional microscopy

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