Ultrahigh resolution multicolor colocalization of single fluorescent nanocrystals

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Description

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 ... continued below

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8 pages

Creation Information

Michalet, X.; Lacoste, T.D.; Pinaud, F.; Chemla, D.S.; Alivisatos, A.P. & Weiss, S. December 20, 2000.

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Description

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.

Physical Description

8 pages

Notes

OSTI as DE00775176

Source

  • BiOS 2001, San Jose, CA (US), 01/20/2001--01/26/2001

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  • Report No.: LBNL--47233
  • Grant Number: AC03-76SF00098
  • Office of Scientific & Technical Information Report Number: 775176
  • Archival Resource Key: ark:/67531/metadc721020

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  • December 20, 2000

Added to The UNT Digital Library

  • Sept. 29, 2015, 5:31 a.m.

Description Last Updated

  • April 5, 2016, 5:41 p.m.

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Citations, Rights, Re-Use

Michalet, X.; Lacoste, T.D.; Pinaud, F.; Chemla, D.S.; Alivisatos, A.P. & Weiss, S. Ultrahigh resolution multicolor colocalization of single fluorescent nanocrystals, article, December 20, 2000; California. (digital.library.unt.edu/ark:/67531/metadc721020/: accessed August 23, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.