Design of multiplex dyes for the detection of different biomolecules

Markus Sauer; Kyung-Tae Han; Volker Ebert; Ralph Mueller; Andreas Schulz; Stefan Seeger; Juergen M. Wolfrum; Jutta Arden-Jacob; Gerhard Deltau; Nicolaas J. Marx; Karl-Heinz Drexhage

doi:10.1117/12.182785

17 August 1994 Design of multiplex dyes for the detection of different biomolecules

Markus Sauer, Kyung-Tae Han, Volker Ebert, Ralph Mueller, Andreas Schulz, Stefan Seeger, Juergen M. Wolfrum, Jutta Arden-Jacob, Gerhard Deltau, Nicolaas J. Marx, Karl-Heinz Drexhage

Author Affiliations +

Proceedings Volume 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV; (1994) https://doi.org/10.1117/12.182785
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

New fluorescent dyes with characteristic fluorescence lifetimes have been developed for bioanalytical applications. Based on the concept of 'multiplex dyes', we have designed several rhodamine dyes with nearly identical absorption and emission spectral characteristics but different fluorescence lifetimes. In order to influence the excited state lifetime without changing the spectral characteristics we modified rhodamines with non- conjugated substituents that promote non-radiative transitions. First investigations with covalently coupled biomolecules show the potential of multiplex dyes in DNA sequencing and antigen detection. The biomolecules are identified through the intrinsic fluorescence lifetime of the dye. This principle offers the possibility to make use of different fluorescence lifetimes in each wavelength region. Therefore the number of discernable tags is greatly enhanced. Extending this principle to applications with laser diodes, new rhodamines with functional groups for covalent coupling of analytes have been developed. The new labels exhibit absorption and emission beyond 600 nm and have a high fluorescence quantum efficiency, even in aqueous buffer systems. Time-resolved and intensity measurements of the dyes covalently coupled to a synthetic oligonucleotide are presented. The results obtained in different capillary systems using laser diodes as excitation sources show the potential of these dyes in the red region of the spectrum.

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Markus Sauer, Kyung-Tae Han, Volker Ebert, Ralph Mueller, Andreas Schulz, Stefan Seeger, Juergen M. Wolfrum, Jutta Arden-Jacob, Gerhard Deltau, Nicolaas J. Marx, and Karl-Heinz Drexhage "Design of multiplex dyes for the detection of different biomolecules", Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); https://doi.org/10.1117/12.182785

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