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Proceedings Article

Ultrahigh-resolution multicolor colocalization of single fluorescent nanocrystals

[+] Author Affiliations
Xavier Michalet, Thilo D. Lacoste, Fabien Pinaud, Daniel S. Chemla, Shimon Weiss

Lawrence Berkeley National Lab. (USA)

A. Paul Alivisatos

Lawrence Berkeley National Lab. and Univ. of California/Berkeley (USA)

Proc. SPIE 4258, Nanoparticles and Nanostructured Surfaces: Novel Reporters with Biological Applications, 8 (June 18, 2001); doi:10.1117/12.430768
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From Conference Volume 4258

  • Nanoparticles and Nanostructured Surfaces: Novel Reporters with Biological Applications
  • Catherine J. Murphy
  • San Jose, CA | January 20, 2001

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.

© (2001) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.
Citation

Xavier Michalet ; Thilo D. Lacoste ; Fabien Pinaud ; Daniel S. Chemla ; A. Paul Alivisatos, et al.
"Ultrahigh-resolution multicolor colocalization of single fluorescent nanocrystals", Proc. SPIE 4258, Nanoparticles and Nanostructured Surfaces: Novel Reporters with Biological Applications, 8 (June 18, 2001); doi:10.1117/12.430768; http://dx.doi.org/10.1117/12.430768


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