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Proceedings Paper

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

The reliability of lanthanide luminescence measurements, by both flow cytometry and digital microscopy, will be enhanced by the availability of narrow-band emitting lanthanide calibration beads. These beads can also be used to characterize spectrographic instruments, including microscopes. Methods: 0.5, 3, and 5 micron (µm) beads containing a luminescent europium-complex were manufactured and the luminescence distribution of the 5 µm beads was measured with a time-delayed luminescence flow cytometer and a timedelayed digital microscope. The distribution of the luminescence intensity from the europium-complex in individual beads was determined on optical sections by confocal microscopy. The emission spectra of the beads under UV excitation were determined with a PARISS® spectrophotometer. The kinetics of the luminescence bleaching caused by UV irradiation were measured under LED excitation with a fluorescence microscope. Results: The kinetics of UV bleaching were very similar for the 0.5, 3, and 5 µm beads. Emission peaks were found at 592, 616, and 685 nanometers (nm). The width of the principal peak at half-maximum (616 nm) was 9.9 nm. The luminescence lifetimes in water and in air were 340 and 460 microseconds (µs), respectively. The distribution of the europium- complex in the beads was homogeneous. Conclusions: The 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: 29 February 2008
PDF: 9 pages
Proc. SPIE 6859, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI, 685917 (29 February 2008); doi: 10.1117/12.762110
Show Author Affiliations
Robert C. Leif, Newport Instruments (United States)
Dayong Jin, Macquarie Univ. (Australia)
James Piper, Macquarie Univ. (Australia)
Lidia M. Vallarino, Virginia Commonwealth Univ. (United States)
John W. Williams, Virginia Commonwealth Univ. (United States)
Sean Yang, Newport Instruments (United States)
Robert M. Zucker, Environmental Protection Agency (United States)

Published in SPIE Proceedings Vol. 6859:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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