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Fast-Gated Intensified Charge-Coupled Device Camera to Record Time-Resolved Fluorescence Spectra of Tryptophan

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Abstract:

The possibilities of a 200 ps gated intensified charge-coupled device (CCD) camera to record time-resolved fluorescence were explored using the fluorescing amino acid tryptophan and its derivative Nacetyl-tryptophan amide (NATA) as model compounds. The results were compared to complementary data from time-correlated single-photon counting (TCSPC) experiments. If a spectral resolution of 1–2 nm is desired, the fast-gated intensified CCD (ICCD) camera is the method of choice. For a 10-5 M tryptophan solution, time-resolved emission spectra and intensity decays (measured over 12 ns at 25 ps resolution) could be obtained in typically 10 minutes, giving the well-known lifetimes of 0.5 and 3 ns. In addition, a longer lifetime of 7 ns was found at the red edge of the spectrum. The very short gate time of the ICCD camera allowed us to observe a shift in the emission maximum of tryptophan even within the first nanosecond of decay of the fluorescence emission. As expected from the tryptophan rotamer model, such a shift is not observed in NATA. Using amplitudes obtained by global analysis, decay-associated spectra of these lifetimes were constructed.

Keywords: CCD; Time-gated charge-coupled device; Time-resolved fluorescence; Tryptophan

Document Type: Research Article

DOI: http://dx.doi.org/10.1366/000370204872908

Affiliations: 1: Department of Analytical Chemistry and Applied Spectroscopy, Vrije Universiteit, Faculty of Sciences, Laser Centre VU, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands 2: Department of Physical Chemistry, Vrije Universiteit, Faculty of Sciences, Laser Centre VU, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands

Publication date: June 1, 2004

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