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Proceedings Paper

Optimizing the luminescence of lanthanide(III) macrocyclic complexes for the detection of anti-5BrdU
Author(s): Robert C. Leif; Margie C. Becker; Alfred J. Bromm; Lidia M. Vallarino; John Williams; Steven A. Williams; Sean Yang
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Paper Abstract

A Eu(III)-macrocycle-mono-isothiocyanate, Quantum Dye®, has been coupled to a monoclonal antibody against 5BrdU. Since Quantum Dyes do not undergo concentration quenching, the coupling conditions were optimized to achieve the maximum number of Eu(III) macrocycles bound to the antiBrdU, without decrease in solubility or loss of antigen-binding ability. In order to optimize the coupling conditions, a colorimetric method for the quantitation of the Eu(III)- macrocycle-mono-isothio-cyanate has been developed. A simple mixture composed of an ethanolic solution and a Gd(III)- containing aqueous solution is now used to provide lanthanide enhanced luminescence, LEL. Under LEL conditions, the specific binding of Eu(III) macrocycles to apoptotic cells has been observed in both aqueous and mounted slide preparations. A comparison between measurements of the same LEL model system, obtained in both time-gated luminescence and standard fluorescence modes, has demonstrated that time- gating significantly improves the signal to noise ratio.

Paper Details

Date Published: 28 May 2002
PDF: 12 pages
Proc. SPIE 4622, Optical Diagnostics of Living Cells V, (28 May 2002); doi: 10.1117/12.468350
Show Author Affiliations
Robert C. Leif, Newport Instruments (United States)
Margie C. Becker, Phoenix Flow Systems (United States)
Alfred J. Bromm, Virginia Commonwealth Univ. (United States)
Lidia M. Vallarino, Virginia Commonwealth Univ. (United States)
John Williams, Virginia Commonwealth Univ. (United States)
Steven A. Williams, Virginia Commonwealth Univ. (United States)
Sean Yang, Newport Instruments (United States)

Published in SPIE Proceedings Vol. 4622:
Optical Diagnostics of Living Cells V
Robert C. Leif; Daniel L. Farkas; Robert C. Leif, Editor(s)

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