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Novel Fluorophores as Building Blocks for Optical Probes for In Vivo Near Infrared Fluorescence (NIRF) Imaging

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Abstract

Aiming at the identification of new fluorescent reporters for targeted optical probes, we assessed the application-relevant features of a novel asymmetric cyanine, DY-681, in comparison to the only clinically approved dye indocyanine green (ICG), the golden imaging standard Cy5.5, and the asymmetric cyanine DY-676 successfully exploited by us for the design of different contrast agents. This comparison included the analysis of the spectroscopic properties of the free fluorophores and their thermal stability in aqueous solution as well as their cytotoxic potential. In addition, the absorption and emission features of IgG-conjugated DY-681 were examined. The trimethine DY-681 exhibited spectral features closely resembling that of the pentamethine Cy5.5. Its high thermal stability in phosphate buffer saline (PBS) solution in conjunction with its low cytotoxicity, reaching similar values as determined for Cy5.5 and DY-676, renders this dye more attractive as ICG and, due to its improved fluorescence quantum yield in PBS, also superior to DY-676. Although in PBS, Cy5.5 was still more fluorescent, the fluorescence quantum yields (Φ f) of DY-681 and Cy5.5 in PBS containing 5 mass-% bovine serum albumin (BSA) were comparable. Labeling experiments with DY-681 and the model antibody IgG revealed promisingly high Φ f values of the bioconjugated dye.

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Acknowledgements

We are grateful to DYOMICS GmbH (Jena, Germany) for the generous supply of the dyes DY-676, DY-681, and DY-682. This work was partially supported by the Thüringer Aufbaubank. Furthermore, we thank Yvonne Heyne for assistance in the cytotoxicity studies and Franziska Hamann for the preparation of the IgG conjugates.

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  1. BAM Federal Institute for Materials Research and Testing, Division I.5, Richard-Willstaetter-Str. 11, 12489, Berlin, Germany

    Jutta Pauli, Robert Brehm, Monika Spieles & Ute Resch-Genger

  2. Institut für Diagnostische und Interventionelle Radiologie des Klinikums der Friedrich-Schiller-Universität Jena (IDIR), Forschungszentrum Lobeda, Erlanger Allee 101, 07747, Jena, Germany

    Werner A. Kaiser & Ingrid Hilger

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  1. Jutta Pauli
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  2. Robert Brehm
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Correspondence to Ingrid Hilger or Ute Resch-Genger.

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Pauli, J., Brehm, R., Spieles, M. et al. Novel Fluorophores as Building Blocks for Optical Probes for In Vivo Near Infrared Fluorescence (NIRF) Imaging. J Fluoresc 20, 681–693 (2010). https://doi.org/10.1007/s10895-010-0603-7

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