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Proceedings Paper

Progress in the use of Quantum Dye Eu(III)-macrocycles
Author(s): Robert C. Leif; Margie C. Becker; Lidia M. Vallarino; John W. Williams; Sean Yang
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Paper Abstract

A Eu(III)-macrocycle-mono-isothiocyanate, Quantum Dye, has been synthesized that has minimal contamination with the Eu(III)-macrocycle-di-isothiocyanate, which cross-links proteins. The mono-isothiocyanate has been conjugated to streptavidin (EuMac-Strept). An indirect assay with EuMac-Strept and biotinylated anti5BrdU has been used to observe apoptotic cells. This system and cells directly labeled with the Eu(III)-macrocycle-di-isothiocyanate have been employed in fading studies and reagent stability tests. The fading of cells mounted in a plastic medium was much slower than that observed when the cells were in the aqueous, micellar Lanthanide Enhanced Luminescence (LEL) solution. The fading was not the result of the photo-destruction of the Eu(III)-macrocycle, since the luminescence returned after a second addition of the LEL solution. A time-gated, peltier cooled, monochrome CCD camera has been combined with a flashlamp to eliminate imaging of the emission of fluorescein while maintaining the images of EuMAc staining. This was demonstrated with both separate preparations of fluorescein and EuMac stained cells and mixtures thereof. Time-gating was employed to produce an EuMac image of cells that were stained with both the EuMac and DAPI.

Paper Details

Date Published: 19 June 2003
PDF: 13 pages
Proc. SPIE 4962, Manipulation and Analysis of Biomolecules, Cells, and Tissues, (19 June 2003); doi: 10.1117/12.486324
Show Author Affiliations
Robert C. Leif, Newport Instruments (United States)
Margie C. Becker, Phoenix Flow Systems (United States)
Lidia M. Vallarino, Virginia Commonwealth Univ. (United States)
John W. Williams, Virginia Commonwealth Univ. (United States)
Sean Yang, Newport Instruments (United States)

Published in SPIE Proceedings Vol. 4962:
Manipulation and Analysis of Biomolecules, Cells, and Tissues
Dan V. Nicolau; Joerg Enderlein; Robert C. Leif; Daniel L. Farkas, Editor(s)

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