Ultrahigh-resolution colocalization of spectrally separable point-like fluorescent probes

Xavier Michalet; Thilo D. Lacoste; Shimon Weiss

doi:10.1006/meth.2001.1218

Ultrahigh-resolution colocalization of spectrally separable point-like fluorescent probes

Xavier Michalet, Thilo D. Lacoste, Shimon Weiss

Lawrence Berkeley National Laboratory

Research output: Contribution to journal › Article › peer-review

53 Scopus citations

Abstract

An ultrahigh-resolution colocalization method based on the simultaneous acquisition and analysis of spectrally separated images of the excitation point-spread function of point-like fluorescent probes is reviewed. It is shown that molecular distances can be measured with accuracy better than 10 nm using conventional far-field optics. A detailed account of the methodology, theoretical considerations, signal processing, and data fitting algorithms is given.

Original language	English
Pages (from-to)	87-102
Number of pages	16
Journal	Methods
Volume	25
Issue number	1
DOIs	https://doi.org/10.1006/meth.2001.1218
State	Published - Sep 2001
Externally published	Yes

Bibliographical note

Funding Information:
We thank Fabien Pinaud and Paul Alivisatos for providing the nanocrystals used in this work; Tsing-Hua Her, Ted Laurence, and Alois Sonnleitner for useful discussions and help with the optics; and Daniel S. Chemla for his continuous and enthusiastic support. This work was supported by the National Institutes of Health, National Center for Research Resources, Grant 1 R01 RR1489101, through the U.S. Department of Energy under Contract DE-AC03-76SF00098. X.M. is an HFSP Postdoctoral Fellow.

Funding

We thank Fabien Pinaud and Paul Alivisatos for providing the nanocrystals used in this work; Tsing-Hua Her, Ted Laurence, and Alois Sonnleitner for useful discussions and help with the optics; and Daniel S. Chemla for his continuous and enthusiastic support. This work was supported by the National Institutes of Health, National Center for Research Resources, Grant 1 R01 RR1489101, through the U.S. Department of Energy under Contract DE-AC03-76SF00098. X.M. is an HFSP Postdoctoral Fellow.

Funders	Funder number
National Institutes of Health
U.S. Department of Energy	DE-AC03-76SF00098
National Center for Research Resources	R01RR014891, 1 R01 RR1489101

Keywords

Confocal
Diffraction limit
Fluorescence
Microscopy
Quantum dot
Semiconductor nanocrystal
Single molecule
Superresolution
TransFluoSphere
Two-photon

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10.1006/meth.2001.1218

Cite this

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abstract = "An ultrahigh-resolution colocalization method based on the simultaneous acquisition and analysis of spectrally separated images of the excitation point-spread function of point-like fluorescent probes is reviewed. It is shown that molecular distances can be measured with accuracy better than 10 nm using conventional far-field optics. A detailed account of the methodology, theoretical considerations, signal processing, and data fitting algorithms is given.",

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Ultrahigh-resolution colocalization of spectrally separable point-like fluorescent probes

Abstract

Bibliographical note

Funding

Keywords

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