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Nanoparticle-encapsulated vis- and NIR-emissive fluorophores with different fluorescence decay kinetics for lifetime multiplexing

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Abstract

Bioanalytical, clinical, and security applications increasingly require simple, efficient, and versatile strategies to measure an ever increasing number of analytes or events in parallel in a broad variety of detection formats as well as in conjunction with chromatographic separation techniques or flow cytometry. An attractive alternative to common optical multiplexing and encoding methods utilizing spectral multiplexing/color encoding and intensity encoding is lifetime multiplexing, which relies on the discrimination between different fluorescent reporters based on their fluorescence decay kinetics. Here, we propose a platform of surface-functionalizable polymeric nanoparticles stained with fluorophores differing in their fluorescence lifetimes as a new multiplexing and encoding approach. Proof-of-concept measurements with different sets of lifetime-encoded polystyrene nanoparticles are presented, obtained via staining of preformed particles with visible (vis)- and near-infrared (NIR)-emissive organic dyes, which display very similar absorption and emission spectra to enable excitation and detection at the same wavelengths, yet sufficiently different fluorescence decay kinetics in suspension, thereby minimizing instrumentation costs. Data analysis was performed with a linear combination approach in the lifetime domain. Our results and first cell experiments with these reporter sets underline the suitability of our multiplexing strategy for the discrimination between and the quantification of different labels. This simple and versatile concept can be extended to all types of fluorophores, thereby expanding the accessible time scale, and can be used, e.g., for the design of labels and targeted probes for fluorescence assays and molecular imaging, cellular imaging studies, and barcoding applications, also in conjunction with spectral and intensity encoding.

Nanoparticle-based lifetime multiplexing in living cells

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Acknowledgments

We gratefully acknowledge the financial support from the Federal Ministry of Economics and Technology (BMWI-22/06 and BMWI-13/09). We would like to thank D. Drescher and J. Kneipp (BAM 1.0) for designing and performing the cell culture experiments. We also express our gratitude to Picoquant GmbH for support with the FLIM setup.

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Authors and Affiliations

  1. BAM Federal Institute for Materials Research and Testing, Richard-Willstaetter-Str. 11, 12489, Berlin, Germany

    Katrin Hoffmann, Thomas Behnke, Markus Grabolle & Ute Resch-Genger

Authors
  1. Katrin Hoffmann
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  2. Thomas Behnke
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  3. Markus Grabolle
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  4. Ute Resch-Genger
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Corresponding author

Correspondence to Ute Resch-Genger.

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Published in the topical collection Multiplex Platforms in Diagnostics and Bioanalytics with guest editors Günter Peine and Günther Proll.

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Hoffmann, K., Behnke, T., Grabolle, M. et al. Nanoparticle-encapsulated vis- and NIR-emissive fluorophores with different fluorescence decay kinetics for lifetime multiplexing. Anal Bioanal Chem 406, 3315–3322 (2014). https://doi.org/10.1007/s00216-013-7597-3

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  • DOI: https://doi.org/10.1007/s00216-013-7597-3

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