Ultrahigh resolution multicolor colocalization of single fluorescent nanocrystals

X. Michalet, T. D. Lacoste, F. Pinaud, D. S. Chemla, A. P. Alivisatos, S. Weiss

Research output: Contribution to journalConference articlepeer-review

4 Scopus citations


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-spreadfunction (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.

Original languageEnglish
Pages (from-to)8-15
Number of pages8
JournalProceedings of SPIE - The International Society for Optical Engineering
StatePublished - 2001
Externally publishedYes
EventNanoparticles and Nanostrutured Surfaces: Novel Reporters with Biological Applications - San Jose, CA, United States
Duration: 24 Jan 200125 Jan 2001


  • Confocal
  • Diffraction limit
  • Fluorescence
  • Microscopy
  • Quantum dot
  • Semiconductor nanocrystal
  • Single molecule
  • Superresolution


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