Ultrahigh-resolution multicolor colocalization of single fluorescent nanocrystals

Xavier Michalet; Thilo D. Lacoste; Fabien Pinaud; Daniel S. Chemla; A. Paul Alivisatos; Shimon Weiss

doi:10.1117/12.430768

18 June 2001 Ultrahigh-resolution multicolor colocalization of single fluorescent nanocrystals

Xavier Michalet, Thilo D. Lacoste, Fabien Pinaud, Daniel S. Chemla, A. Paul Alivisatos, Shimon Weiss

Proceedings Volume 4258, Nanoparticles and Nanostructured Surfaces: Novel Reporters with Biological Applications; (2001) https://doi.org/10.1117/12.430768
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

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.

Citation Download Citation

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

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