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Proceedings Paper

Improved molecular barcodes by lifetime discrimination
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Paper Abstract

Individual microspheres labeled with a unique barcode and a surface-bound probe are able to provide multiplexed biological assays in a convenient and high-throughput format. Typically, barcodes are created by impregnating microspheres with several colors of fluorophores mixed at different intensity levels. The number of barcodes is limited to hundreds primarily due to variability in fluorophore loading and difficulties in compensating for signal crosstalk. We constructed a molecular barcode based on differences in lifetimes rather than intensities. Lifetime-based measurements have an advantage in that signal from neighboring channels is reduced (because signal intensities are equal) and may be mathematically deconvoluted. The excited state lifetime of quantum dots (QDs) was systematically altered by attaching a variable number of quencher molecules to the surface. We have synthesized a series of ten QDs with distinguishable lifetimes all emitting at the same wavelength. The QDs were loaded into microspheres to determine the expected signal intensities. The uncertainty in lifetimes as a function of the interrogation time was determined. An acceptable standard deviation (3%) was obtained with a measurement time of approximately 10-30 μsec. Currently, we are expanding these studies to include multiple wavelengths and determining the maximal number of barcodes for a given spectral window.

Paper Details

Date Published: 3 March 2009
PDF: 12 pages
Proc. SPIE 7189, Colloidal Quantum Dots for Biomedical Applications IV, 71890R (3 March 2009); doi: 10.1117/12.809400
Show Author Affiliations
Daniel B. Hall, Radiation Monitoring Devices, Inc. (United States)
William G. Lawrence, Radiation Monitoring Devices, Inc. (United States)

Published in SPIE Proceedings Vol. 7189:
Colloidal Quantum Dots for Biomedical Applications IV
Marek Osinski; Thomas M. Jovin M.D.; Kenji Yamamoto M.D., Editor(s)

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