Time-resolved fluorescence measurements using stroboscopic excitation

Daniel R. Matthews; Huw D. Summers; Kerenza Njoh; Rachel J. Errington; Paul J. Smith; Simon Ameer-Beg; Boris Vojnovic

doi:10.1117/12.632914

1 September 2005 Time-resolved fluorescence measurements using stroboscopic excitation

Daniel R. Matthews, Huw D. Summers, Kerenza Njoh, Rachel J. Errington, Paul J. Smith, Simon Ameer-Beg, Boris Vojnovic

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Proceedings Volume 5864, Novel Optical Instrumentation for Biomedical Applications II; 586409 (2005) https://doi.org/10.1117/12.632914
Event: European Conferences on Biomedical Optics 2005, 2005, Munich, Germany

Abstract

We report on the development of a simple technique for obtaining time-domain information using dc detection of fluorescence. We show that this is feasible for assays where a change in lifetime of an indicator occurs in reaction to an analyte, in fluorescence resonance energy transfer for example, and could be particularly useful for assays performed in the scaled-down environment of a "lab-on-a-chip". A rate equation model is presented which allows an objective analysis of the relative importance of the key measurement parameters: optical saturation of the fluorophore and excitation pulse characteristics. We present a comparison of the model with a cuvette based analysis of a carbocyanine dye where the excitation source is a 650 nm wavelength, self-pulsing AlGaInP laser diode.

Citation Download Citation

Daniel R. Matthews, Huw D. Summers, Kerenza Njoh, Rachel J. Errington, Paul J. Smith, Simon Ameer-Beg, and Boris Vojnovic "Time-resolved fluorescence measurements using stroboscopic excitation", Proc. SPIE 5864, Novel Optical Instrumentation for Biomedical Applications II, 586409 (1 September 2005); https://doi.org/10.1117/12.632914

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