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Proceedings Paper

Ultrahigh-resolution multicolor colocalization of single fluorescent nanocrystals
Author(s): Xavier Michalet; Thilo D. Lacoste; Fabien Pinaud; Daniel S. Chemla; A. Paul Alivisatos; Shimon Weiss
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Paper 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.

Paper Details

Date Published: 18 June 2001
PDF: 8 pages
Proc. SPIE 4258, Nanoparticles and Nanostructured Surfaces: Novel Reporters with Biological Applications, (18 June 2001); doi: 10.1117/12.430768
Show Author Affiliations
Xavier Michalet, Lawrence Berkeley National Lab. (United States)
Thilo D. Lacoste, Lawrence Berkeley National Lab. (United States)
Fabien Pinaud, Lawrence Berkeley National Lab. (United States)
Daniel S. Chemla, Lawrence Berkeley National Lab. (United States)
A. Paul Alivisatos, Lawrence Berkeley National Lab. and Univ. of California/Berkeley (United States)
Shimon Weiss, Lawrence Berkeley National Lab. (United States)


Published in SPIE Proceedings Vol. 4258:
Nanoparticles and Nanostructured Surfaces: Novel Reporters with Biological Applications
Catherine J. Murphy, Editor(s)

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