Improved molecular barcodes by lifetime discrimination

Daniel B. Hall; William G. Lawrence

doi:10.1117/12.809400

3 March 2009 Improved molecular barcodes by lifetime discrimination

Daniel B. Hall, William G. Lawrence

Proceedings Volume 7189, Colloidal Quantum Dots for Biomedical Applications IV; 71890R (2009) https://doi.org/10.1117/12.809400
Event: SPIE BiOS, 2009, San Jose, California, United States

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.

Citation Download Citation

Daniel B. Hall and William G. Lawrence "Improved molecular barcodes by lifetime discrimination", Proc. SPIE 7189, Colloidal Quantum Dots for Biomedical Applications IV, 71890R (3 March 2009); https://doi.org/10.1117/12.809400

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KEYWORDS

Luminescence

Quantum dots

Molecules

Pulsed laser operation

Absorption

Oscilloscopes

Silica

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