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. 2008 Oct;34(5):487-93.
doi: 10.1007/s10867-008-9107-y. Epub 2008 Aug 27.

Triple-FRET Technique for Energy Transfer Between Conjugated Polymer and TAMRA Dye with Possible Applications in Medical Diagnostics

Anamika Aneja  1 , Nidhi Mathur, P K Bhatnagar, P C Mathur
Affiliations

Triple-FRET Technique for Energy Transfer Between Conjugated Polymer and TAMRA Dye with Possible Applications in Medical Diagnostics

Anamika Aneja et al. J Biol Phys. 2008 Oct.

Abstract

Three-component Förster resonance energy transfer (FRET) has been used to obtain efficient FRET between the cationic conjugated polymer (CCP) as donor and 5-carboxy tetramethylrhodamine (TAMRA) dye as acceptor, by using an intermediate donor, fluorescein. In spite of the fact that there is enough overlap between the emission spectra of CCP and absorption spectra of TAMRA, the efficiency of FRET between CCP and TAMRA is poor. The reason for this is that while the Förster critical distance is not very sensitive to the overlap, the FRET efficiency is extremely sensitive to it. However, it is observed that the FRET efficiency between CCP and TAMRA improves considerably when fluorescein is introduced in the solution. The triple FRET so obtained can be used for deoxyribonucleic acid sequence detection in medical diagnostics because the fluorescence emission from TAMRA is pH-insensitive.

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Figures

Fig. 1
Fig. 1
Absorption and emission spectra of the light-emitting polymer and fluorescent dyes—PFP: A1 (λmax) = 376 nm; F1 (λmax) = 410 nm; fluorescein: A2 (λmax) = 492 nm, F2 (λmax) = 520 nm; TAMRA: A3 (λmax) = 547 nm, F3 (λ max ) = 573 nm
Fig. 2
Fig. 2
Emission spectra of polymer PFP, fluorescein (FL), and TAMRA in a composite solution for different concentrations of PFP (from 0 to 20 μl) at an excitation of 376 nm
Fig. 3
Fig. 3
Comparison of the emission spectra of conventional FRET (PFP/TAMRA, crosses) and two-step FRET (PFP/FL/TAMRA, open squares)
Fig. 4
Fig. 4
Effect of fluorescein on the PL intensity of TAMRA for different concentrations of CCP
See this image and copyright information in PMC

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