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Practical Time-Resolved Fluorescence Spectroscopy: Avoiding Artifacts and Using Lifetime Standards

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Standardization and Quality Assurance in Fluorescence Measurements I

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 5))

Abstract

In this chapter we describe how artifacts can be avoided in the two most commonly used time-resolved fluorometries, namely the single-photon timing and the multifrequency phase-modulation techniques. The most frequently encountered artifacts (inner filter effect, autofluorescence, polarization effects, color effect, photobleaching, deoxygenation, pulse pile-up, and linearity of the time response in the time-to-amplitude converter) are described in detail and remedies are presented to avoid these pitfalls. An extensive list of fluorescence lifetime standards is presented, which allows the spectroscopist to calibrate and test time-resolved instruments for systematic errors.

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Author information

Authors and Affiliations

  1. Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200f, bus 02404, 3001, Heverlee, Belgium

    Noël Boens

  2. BIOMED, Universiteit Hasselt, Agoralaan, Building D, 3590, Diepenbeek, Belgium

    Marcel Ameloot

  3. CNRS UMR 8531, Laboratoire de Chimie Générale, CNAM, 292 rue Saint-Martin, 75141, Paris Cedex 03, France

    Bernard Valeur

  4. Laboratoire PPSM, ENS-Cachan, 61 avenue du Président Wilson, 94235, Cachan Cedex, France

    Bernard Valeur

Authors
  1. Noël Boens
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  2. Marcel Ameloot
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  3. Bernard Valeur
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Corresponding author

Correspondence to Noël Boens .

Editor information

Ute Resch-Genger

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© 2008 Springer-Verlag Berlin Heidelberg

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Boens, N., Ameloot, M., Valeur, B. (2008). Practical Time-Resolved Fluorescence Spectroscopy: Avoiding Artifacts and Using Lifetime Standards. In: Resch-Genger, U. (eds) Standardization and Quality Assurance in Fluorescence Measurements I. Springer Series on Fluorescence, vol 5. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2008_044

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