Proposal of a New Method for Measuring Förster Resonance Energy Transfer (FRET) Rapidly, Quantitatively and Non-Destructively
The process of radiationless energy transfer from a chromophore in an excited electronic state (the “donor”) to another chromophore (an “acceptor”), in which the energy released by the donor effects an electronic transition, is known as “Förster Resonance Energy Transfer” (FRET). The rate of energy...
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| Format: | Article |
| Language: | English |
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MDPI AG
2012-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | http://www.mdpi.com/1422-0067/13/10/12367 |
| _version_ | 1811338814903287808 |
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| author | Paul Johannes Helm |
| author_facet | Paul Johannes Helm |
| author_sort | Paul Johannes Helm |
| collection | DOAJ |
| description | The process of radiationless energy transfer from a chromophore in an excited electronic state (the “donor”) to another chromophore (an “acceptor”), in which the energy released by the donor effects an electronic transition, is known as “Förster Resonance Energy Transfer” (FRET). The rate of energy transfer is dependent on the sixth power of the distance between donor and acceptor. Determining FRET efficiencies is tantamount to measuring distances between molecules. A new method is proposed for determining FRET efficiencies rapidly, quantitatively, and non-destructively on ensembles containing donor acceptor pairs: at wavelengths suitable for mutually exclusive excitations of donors and acceptors, two laser beams are intensity-modulated in rectangular patterns at duty cycle ½ and frequencies ƒ1 and ƒ2 by electro-optic modulators. In an ensemble exposed to these laser beams, the donor excitation is modulated at ƒ1, and the acceptor excitation, and therefore the degree of saturation of the excited electronic state of the acceptors, is modulated at ƒ2. Since the ensemble contains donor acceptor pairs engaged in FRET, the released donor fluorescence is modulated not only at ƒ1 but also at the beat frequency Δƒ: = |ƒ1 − ƒ2|. The depth of the latter modulation, detectable via a lock-in amplifier, quantitatively indicates the FRET efficiency. |
| first_indexed | 2024-04-13T18:16:06Z |
| format | Article |
| id | doaj.art-7eb16dd3f7e14d10aafc8398f3932e6c |
| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-13T18:16:06Z |
| publishDate | 2012-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-7eb16dd3f7e14d10aafc8398f3932e6c2022-12-22T02:35:40ZengMDPI AGInternational Journal of Molecular Sciences1422-00672012-09-011310123671238210.3390/ijms131012367Proposal of a New Method for Measuring Förster Resonance Energy Transfer (FRET) Rapidly, Quantitatively and Non-DestructivelyPaul Johannes HelmThe process of radiationless energy transfer from a chromophore in an excited electronic state (the “donor”) to another chromophore (an “acceptor”), in which the energy released by the donor effects an electronic transition, is known as “Förster Resonance Energy Transfer” (FRET). The rate of energy transfer is dependent on the sixth power of the distance between donor and acceptor. Determining FRET efficiencies is tantamount to measuring distances between molecules. A new method is proposed for determining FRET efficiencies rapidly, quantitatively, and non-destructively on ensembles containing donor acceptor pairs: at wavelengths suitable for mutually exclusive excitations of donors and acceptors, two laser beams are intensity-modulated in rectangular patterns at duty cycle ½ and frequencies ƒ1 and ƒ2 by electro-optic modulators. In an ensemble exposed to these laser beams, the donor excitation is modulated at ƒ1, and the acceptor excitation, and therefore the degree of saturation of the excited electronic state of the acceptors, is modulated at ƒ2. Since the ensemble contains donor acceptor pairs engaged in FRET, the released donor fluorescence is modulated not only at ƒ1 but also at the beat frequency Δƒ: = |ƒ1 − ƒ2|. The depth of the latter modulation, detectable via a lock-in amplifier, quantitatively indicates the FRET efficiency.http://www.mdpi.com/1422-0067/13/10/12367FRETLSMdynamicnon-destructiveelectro-optic modulatorbeatsaturationmodulation |
| spellingShingle | Paul Johannes Helm Proposal of a New Method for Measuring Förster Resonance Energy Transfer (FRET) Rapidly, Quantitatively and Non-Destructively International Journal of Molecular Sciences FRET LSM dynamic non-destructive electro-optic modulator beat saturation modulation |
| title | Proposal of a New Method for Measuring Förster Resonance Energy Transfer (FRET) Rapidly, Quantitatively and Non-Destructively |
| title_full | Proposal of a New Method for Measuring Förster Resonance Energy Transfer (FRET) Rapidly, Quantitatively and Non-Destructively |
| title_fullStr | Proposal of a New Method for Measuring Förster Resonance Energy Transfer (FRET) Rapidly, Quantitatively and Non-Destructively |
| title_full_unstemmed | Proposal of a New Method for Measuring Förster Resonance Energy Transfer (FRET) Rapidly, Quantitatively and Non-Destructively |
| title_short | Proposal of a New Method for Measuring Förster Resonance Energy Transfer (FRET) Rapidly, Quantitatively and Non-Destructively |
| title_sort | proposal of a new method for measuring forster resonance energy transfer fret rapidly quantitatively and non destructively |
| topic | FRET LSM dynamic non-destructive electro-optic modulator beat saturation modulation |
| url | http://www.mdpi.com/1422-0067/13/10/12367 |
| work_keys_str_mv | AT pauljohanneshelm proposalofanewmethodformeasuringforsterresonanceenergytransferfretrapidlyquantitativelyandnondestructively |