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

56 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

Access to Document

10.1006/meth.2001.1218

Cite this

@article{8775bf4021b3491fa440f2020d09d1ab,

title = "Ultrahigh-resolution colocalization of spectrally separable point-like fluorescent probes",

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.",

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

author = "Xavier Michalet and Lacoste, \{Thilo D.\} and Shimon Weiss",

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.",

year = "2001",

month = sep,

doi = "10.1006/meth.2001.1218",

language = "אנגלית",

volume = "25",

pages = "87--102",

journal = "Methods",

issn = "1046-2023",

publisher = "Academic Press Inc.",

number = "1",

}

TY - JOUR

T1 - Ultrahigh-resolution colocalization of spectrally separable point-like fluorescent probes

AU - Michalet, Xavier

AU - Lacoste, Thilo D.

AU - Weiss, Shimon

N1 - 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.

PY - 2001/9

Y1 - 2001/9

N2 - 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.

AB - 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.

KW - Confocal

KW - Diffraction limit

KW - Fluorescence

KW - Microscopy

KW - Quantum dot

KW - Semiconductor nanocrystal

KW - Single molecule

KW - Superresolution

KW - TransFluoSphere

KW - Two-photon

UR - https://www.scopus.com/pages/publications/0034807907

U2 - 10.1006/meth.2001.1218

DO - 10.1006/meth.2001.1218

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

C2 - 11559000

AN - SCOPUS:0034807907

SN - 1046-2023

VL - 25

SP - 87

EP - 102

JO - Methods

JF - Methods

IS - 1

ER -

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

Abstract

Bibliographical note

Funding

Keywords

Access to Document

Other files and links

Fingerprint

Cite this