Demonstration of FRET in solutions

Sunil Shah; Zygmunt Gryczynski; Rahul Chib; Rafal Fudala; Aatmun Baxi; Julian Borejdo; Anna Synak; Ignacy Gryczynski

doi:10.1088/2050-6120/4/1/015001

Demonstration of FRET in solutions

Sunil Shah, Zygmunt Gryczynski, Rahul Chib, Rafal Fudala, Aatmun Baxi, Julian Borejdo, Anna Synak, Ignacy Gryczynski

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2 Scopus citations

Abstract

We measured the Förster resonance energy transfer (FRET) from Uranin (U) donor to Rhodamine 101 (R101) acceptor in propylene glycol. Steady-state fluorescence measurements show a significant difference between mixed and unmixed fluorophore solutions. In the solution with mixed fluorophores, fluorescence intensity of the U donor decreases and intensity of R101 fluorescence increases. This is visualized as a color change from green to orange. Fluorescence anisotropy of the mixture solution increases in the donor emission wavelength region and decreases in the acceptor emission wavelengths; which is consistent with FRET occurrence. Time-resolved (lifetime) measurements show a decrease of the U lifetime in the presence of R101 acceptor. In the intensity decay of R101 acceptor appears a negative component indicating excited state process. All these measurements prove the presence of FRET in U/R101 mixture fluorescence.

Original language	English
Article number	015001
Journal	Methods and Applications in Fluorescence
Volume	4
Issue number	1
DOIs	https://doi.org/10.1088/2050-6120/4/1/015001
State	Published - Mar 2016

Keywords

3D FRET
FRET in solution
Fluorescence
Forster distance

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title = "Demonstration of FRET in solutions",

abstract = "We measured the F{\"o}rster resonance energy transfer (FRET) from Uranin (U) donor to Rhodamine 101 (R101) acceptor in propylene glycol. Steady-state fluorescence measurements show a significant difference between mixed and unmixed fluorophore solutions. In the solution with mixed fluorophores, fluorescence intensity of the U donor decreases and intensity of R101 fluorescence increases. This is visualized as a color change from green to orange. Fluorescence anisotropy of the mixture solution increases in the donor emission wavelength region and decreases in the acceptor emission wavelengths; which is consistent with FRET occurrence. Time-resolved (lifetime) measurements show a decrease of the U lifetime in the presence of R101 acceptor. In the intensity decay of R101 acceptor appears a negative component indicating excited state process. All these measurements prove the presence of FRET in U/R101 mixture fluorescence.",

keywords = "3D FRET, FRET in solution, Fluorescence, Forster distance",

author = "Sunil Shah and Zygmunt Gryczynski and Rahul Chib and Rafal Fudala and Aatmun Baxi and Julian Borejdo and Anna Synak and Ignacy Gryczynski",

note = "Funding Information: The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This work was supported by the NIH grant R01EB12003 (Z G), and NSF grant CBET-1264608 (I G). A S acknowledges support from National Science Center (Poland) 2015/17/B/ST5/03143. We thank Dr Rafal Luchowski for the calculation of R0 value. Competing interests: The authors have declared that no competing interests exist. We dedicate this paper to Dr J?zef Ku?ba on theoccasion of his 70th birthday. Publisher Copyright: {\textcopyright} 2016 IOP Publishing Ltd.",

year = "2016",

month = mar,

doi = "10.1088/2050-6120/4/1/015001",

language = "English",

volume = "4",

journal = "Methods and applications in fluorescence",

issn = "2050-6120",

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AU - Shah, Sunil

AU - Gryczynski, Zygmunt

AU - Chib, Rahul

AU - Fudala, Rafal

AU - Baxi, Aatmun

AU - Borejdo, Julian

AU - Synak, Anna

AU - Gryczynski, Ignacy

N1 - Funding Information: The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This work was supported by the NIH grant R01EB12003 (Z G), and NSF grant CBET-1264608 (I G). A S acknowledges support from National Science Center (Poland) 2015/17/B/ST5/03143. We thank Dr Rafal Luchowski for the calculation of R0 value. Competing interests: The authors have declared that no competing interests exist. We dedicate this paper to Dr J?zef Ku?ba on theoccasion of his 70th birthday. Publisher Copyright: © 2016 IOP Publishing Ltd.

PY - 2016/3

Y1 - 2016/3

N2 - We measured the Förster resonance energy transfer (FRET) from Uranin (U) donor to Rhodamine 101 (R101) acceptor in propylene glycol. Steady-state fluorescence measurements show a significant difference between mixed and unmixed fluorophore solutions. In the solution with mixed fluorophores, fluorescence intensity of the U donor decreases and intensity of R101 fluorescence increases. This is visualized as a color change from green to orange. Fluorescence anisotropy of the mixture solution increases in the donor emission wavelength region and decreases in the acceptor emission wavelengths; which is consistent with FRET occurrence. Time-resolved (lifetime) measurements show a decrease of the U lifetime in the presence of R101 acceptor. In the intensity decay of R101 acceptor appears a negative component indicating excited state process. All these measurements prove the presence of FRET in U/R101 mixture fluorescence.

AB - We measured the Förster resonance energy transfer (FRET) from Uranin (U) donor to Rhodamine 101 (R101) acceptor in propylene glycol. Steady-state fluorescence measurements show a significant difference between mixed and unmixed fluorophore solutions. In the solution with mixed fluorophores, fluorescence intensity of the U donor decreases and intensity of R101 fluorescence increases. This is visualized as a color change from green to orange. Fluorescence anisotropy of the mixture solution increases in the donor emission wavelength region and decreases in the acceptor emission wavelengths; which is consistent with FRET occurrence. Time-resolved (lifetime) measurements show a decrease of the U lifetime in the presence of R101 acceptor. In the intensity decay of R101 acceptor appears a negative component indicating excited state process. All these measurements prove the presence of FRET in U/R101 mixture fluorescence.

KW - 3D FRET

KW - FRET in solution

KW - Fluorescence

KW - Forster distance

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DO - 10.1088/2050-6120/4/1/015001

M3 - Article

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JO - Methods and applications in fluorescence

JF - Methods and applications in fluorescence

SN - 2050-6120

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Demonstration of FRET in solutions

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