Ultrahigh resolution multicolor colocalization of single fluorescent nanocrystals
Skip to main content
eScholarship
Open Access Publications from the University of California

Ultrahigh resolution multicolor colocalization of single fluorescent nanocrystals

  • Author(s): Michalet, X.
  • Lacoste, T.D.
  • Pinaud, F.
  • Chemla, D.S.
  • Alivisatos, A.P.
  • Weiss, S.
  • et al.
Abstract

A new method for in vitro and possibly in vivo ultrahigh-resolution colocalization and distance measurement between biomolecules is described, based on semiconductor nanocrystal probes. This ruler bridges the gap between FRET and far-field (or near-field scanning optical microscope) imaging and has a dynamic range from few nanometers to tens of micrometers. The ruler is based on a stage-scanning confocal microscope that allows the simultaneous excitation and localization of the excitation point-spread-function (PSF) of various colors nanocrystals while maintaining perfect registry between the channels. Fit of the observed diffraction and photophysics-limited images of the PSFs with a two-dimensional Gaussian allows one to determine their position with nanometer accuracy. This new high-resolution tool opens new windows in various molecular, cell biology and biotechnology applications.

Main Content
Current View