Calibration beads containing luminescent lanthanide ion complexes

Robert C. Leif*, Sean Yang, Dayong Jin, James Piper, Lidia M. Vallarino, John W. Williams, Robert M. Zucker

*Corresponding author for this work

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The reliability of lanthanide luminescence measurements, by both flow cytometry and digital microscopy, would be enhanced by the availability of narrowband emitting, UV excited lanthanide calibration beads. 0.5-, 3-, and 5-μm beads containing a luminescent europium-complex are manufactured. The luminescence distribution of the 5-μm beads is measured with a time-delayed light-scatter-gated luminescence flow cytometer to have a 7.0% coefficient of variation (CV) The spacial distribution of the europium-complex in individual beads is determined to be homogeneous by confocal microscopy. Emission peaks are found at 592, 616 (width 9.9nm), and 685nm with a PARISS® spectrophotometer. The kinetics of the luminescence bleaching caused by UV irradiation of the 0.5- and 5-μm beads measured under LED excitation with a fluorescence microscope indicate that bleaching does not interfere with their imaging. The luminescence lifetimes in water and air were 340 and 460μs, respectively. Thus, these 5-μm beads can be used for spectral calibration of microscopes equipped with a spectrograph, as test particles for time-delayed luminescence flow cytometers, and possibly as labels for macromolecules and cells.

Original languageEnglish
Article number024022
Pages (from-to)1-7
Number of pages7
JournalJournal of Biomedical Optics
Issue number2
Publication statusPublished - 2009

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    Leif, R. C., Yang, S., Jin, D., Piper, J., Vallarino, L. M., Williams, J. W., & Zucker, R. M. (2009). Calibration beads containing luminescent lanthanide ion complexes. Journal of Biomedical Optics, 14(2), 1-7. [024022].