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Journal of Biomedical Optics

Calibration beads containing luminescent lanthanide ion complexes
Author(s): Robert C. Leif; Sean Yang; Dayong Jin; James A. Piper; Lidia M. Vallarino; John W. Williams; Robert M. Zucker
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Paper Abstract

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 9.9 nm), and 685 nm 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 460 μs, 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.

Paper Details

Date Published: 1 March 2009
PDF: 7 pages
J. Biomed. Opt. 14(2) 024022 doi: 10.1117/1.3103646
Published in: Journal of Biomedical Optics Volume 14, Issue 2
Show Author Affiliations
Robert C. Leif, Newport Instruments (United States)
Sean Yang, Newport Instruments (United States)
Dayong Jin, Macquarie Univ. (Australia)
James A. Piper, Macquarie Univ. (Australia)
Lidia M. Vallarino, Virginia Commonwealth Univ. (United States)
John W. Williams, Virginia Commonwealth Univ. (United States)
Robert M. Zucker, U.S. Environmental Protection Agency (United States)

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