Periodic acceptor excitation spectroscopy of single molecules

Sören Doose; Mike Heilemann; Xavier Michalet; Shimon Weiss; Achillefs N. Kapanidis

doi:10.1007/s00249-007-0133-7

Periodic acceptor excitation spectroscopy of single molecules

Sören Doose, Mike Heilemann, Xavier Michalet, Shimon Weiss, Achillefs N. Kapanidis

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

18 Scopus citations

Abstract

Alternating-laser excitation (ALEX) spectroscopy has recently been added to the single-molecule spectroscopy toolkit. ALEX monitors interaction and stoichiometry of biomolecules, reports on biomolecular structure by measuring accurate Förster resonance energy transfer (FRET) efficiencies, and allows sorting of subpopulations on the basis of stoichiometry and FRET. Here, we demonstrate that a simple combination of one continuous-wave donor-excitation laser and one directly modulated acceptor-excitation laser (Periodic Acceptor eXcitation) is sufficient to recapitulate the capabilities of ALEX while minimizing the cost and complexity associated with use of modulation techniques.

Original language	English
Pages (from-to)	669-674
Number of pages	6
Journal	European Biophysics Journal
Volume	36
Issue number	6
DOIs	https://doi.org/10.1007/s00249-007-0133-7
State	Published - Jul 2007
Externally published	Yes

Bibliographical note

Funding Information:
Acknowledgments We thank N.K. Lee for providing DNA samples and L. LeReste for assistance. This work was funded by NIH grant GM069709-01 to S.W. and A.N.K., DOE grants 02ER63339 and 04ER63938 to S.W., and EPSRC grant EP/ D058775, EU Marie Curie Fellowship MIRG-CT-2005-031079, and a UK Bionanotechnology IRC grant to A.N.K. M.H. was supported by a DAAD fellowship.

Funding

Acknowledgments We thank N.K. Lee for providing DNA samples and L. LeReste for assistance. This work was funded by NIH grant GM069709-01 to S.W. and A.N.K., DOE grants 02ER63339 and 04ER63938 to S.W., and EPSRC grant EP/ D058775, EU Marie Curie Fellowship MIRG-CT-2005-031079, and a UK Bionanotechnology IRC grant to A.N.K. M.H. was supported by a DAAD fellowship.

Funders	Funder number
National Institutes of Health
U.S. Department of Energy	02ER63339, 04ER63938
National Institute of General Medical Sciences	R01GM069709
Engineering and Physical Sciences Research Council	EP/ D058775
European Commission	MIRG-CT-2005-031079
Deutscher Akademischer Austauschdienst

Keywords

Alternating-laser excitation (ALEX)
Biomolecular interactions
Förster resonance energy transfer (FRET)
Single-molecule fluorescence spectroscopy

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10.1007/s00249-007-0133-7

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title = "Periodic acceptor excitation spectroscopy of single molecules",

abstract = "Alternating-laser excitation (ALEX) spectroscopy has recently been added to the single-molecule spectroscopy toolkit. ALEX monitors interaction and stoichiometry of biomolecules, reports on biomolecular structure by measuring accurate F{\"o}rster resonance energy transfer (FRET) efficiencies, and allows sorting of subpopulations on the basis of stoichiometry and FRET. Here, we demonstrate that a simple combination of one continuous-wave donor-excitation laser and one directly modulated acceptor-excitation laser (Periodic Acceptor eXcitation) is sufficient to recapitulate the capabilities of ALEX while minimizing the cost and complexity associated with use of modulation techniques.",

keywords = "Alternating-laser excitation (ALEX), Biomolecular interactions, F{\"o}rster resonance energy transfer (FRET), Single-molecule fluorescence spectroscopy",

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note = "Funding Information: Acknowledgments We thank N.K. Lee for providing DNA samples and L. LeReste for assistance. This work was funded by NIH grant GM069709-01 to S.W. and A.N.K., DOE grants 02ER63339 and 04ER63938 to S.W., and EPSRC grant EP/ D058775, EU Marie Curie Fellowship MIRG-CT-2005-031079, and a UK Bionanotechnology IRC grant to A.N.K. M.H. was supported by a DAAD fellowship.",

year = "2007",

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doi = "10.1007/s00249-007-0133-7",

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AU - Heilemann, Mike

AU - Michalet, Xavier

AU - Weiss, Shimon

AU - Kapanidis, Achillefs N.

N1 - Funding Information: Acknowledgments We thank N.K. Lee for providing DNA samples and L. LeReste for assistance. This work was funded by NIH grant GM069709-01 to S.W. and A.N.K., DOE grants 02ER63339 and 04ER63938 to S.W., and EPSRC grant EP/ D058775, EU Marie Curie Fellowship MIRG-CT-2005-031079, and a UK Bionanotechnology IRC grant to A.N.K. M.H. was supported by a DAAD fellowship.

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AB - Alternating-laser excitation (ALEX) spectroscopy has recently been added to the single-molecule spectroscopy toolkit. ALEX monitors interaction and stoichiometry of biomolecules, reports on biomolecular structure by measuring accurate Förster resonance energy transfer (FRET) efficiencies, and allows sorting of subpopulations on the basis of stoichiometry and FRET. Here, we demonstrate that a simple combination of one continuous-wave donor-excitation laser and one directly modulated acceptor-excitation laser (Periodic Acceptor eXcitation) is sufficient to recapitulate the capabilities of ALEX while minimizing the cost and complexity associated with use of modulation techniques.

KW - Alternating-laser excitation (ALEX)

KW - Biomolecular interactions

KW - Förster resonance energy transfer (FRET)

KW - Single-molecule fluorescence spectroscopy

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Periodic acceptor excitation spectroscopy of single molecules

Abstract

Bibliographical note

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Keywords

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