Luminescence energy transfer using a terbium chelate: Improvements on fluorescence energy transfer

Journal Article · Tue Oct 11 04:00:00 EDT 1994 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.91.21.10024· OSTI ID:86528

Selvin, P R; Hearst, J E ^[1]

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1. Lawrence Berkeley Lab., CA (United States)

The authors extend the technique of fluorescence resonance energy transfer (FRET) by introducing a luminescent terbium chelate as a donor and an organic dye, tetramethylrhodamine, as an acceptor. The results are consistent with a Foerster theory of energy transfer, provided the appropriate parameters are used. The use of lanthanide donors in general, and this pair in particular, has many advantages over more conventional FRET pairs, which rely solely on organic dyes. The distance at which 50% energy transfer occurs is large, 65 {angstrom}; the donor lifetime is a single exponential and long (millisecond), making lifetime measurements facile and accurate. Uncertainty in the orientation factor, which creates uncertainty in measured distances, is minimized by the donor`s multiple electronic transitions and long lifetime. The sensitized emission of the acceptor can be measured with little or no interfering background, yielding a >25-fold improvement in the signal-to-background ratio over standard donor-acceptor pairs. These improvements are expected to make distances >100 {angstrom} measurable via FRET. The authors also report measurement of the sensitized emission lifetime, a measurement that is completely insensitive to total concentration and incomplete labeling.

• Sponsoring Organization: USDOE
• DOE Contract Number: AC03-76SF00098
• OSTI ID: 86528
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 21 Vol. 91; ISSN PNASA6; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

40 CHEMISTRY
ELECTRIC DIPOLES
ENERGY TRANSFER
FLUORESCENCE SPECTROSCOPY
INTERATOMIC DISTANCES
RESONANCE FLUORESCENCE
RHODAMINES
TERBIUM COMPLEXES