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New Trends in Fluorescence Spectroscopy pp 277–296Cite as

Two-Photon Fluorescence Fluctuation Spectroscopy

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Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 1))

Abstract

Two-photon excitation spectroscopy has inherent 3-D resolution with excitation volumes as small as 0.1 fl. Compared to conventional fluorometers a reduction by a factor of 1010 of the excitation volume can be achieved The fluorescence fluctuations within the small excitation volume provide, via fluorescence correlation spectroscopy (FCS), a unique way to study biological phenomena In this contribution, we outline the instrumentation of two-photon fluorescence correlation spectroscopy and highlight technical details of our experimental setup We discuss the autocorrelation analysis for single and multiple species with emphasis on the normalized fluctuation amplitude G(0) Furthermore, we revisit another data analysis technique called moment analysis This method was originally introduced 10 years ago and provides a very fast and convenient characterization of the fluctuation amplitude We compare experimental results from moment analysis with that of another data analysis method, the photon counting histogram (PCH), and discussed differences between these two methods.

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Authors
  1. Y. Chen
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  2. J. D. Müller
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  3. J. S. Eid
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  4. E. Gratton
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Editor information

Editors and Affiliations

  1. Conservatoire National des Arts et Métiers, Laboratoire de Chimie Général, 292 rue Saint-Martin, 75141, Paris Cedex 03, France

    Bernard Valeur

  2. Ecole Normale Supérieure de Cachan, 61 Av. du Président Wilson, 94235, Cachan Cedex, France

    Jean-Claude Brochon

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© 2001 Springer-Verlag Berlin Heidelberg

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Chen, Y., Müller, J.D., Eid, J.S., Gratton, E. (2001). Two-Photon Fluorescence Fluctuation Spectroscopy. In: Valeur, B., Brochon, JC. (eds) New Trends in Fluorescence Spectroscopy. Springer Series on Fluorescence, vol 1. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-56853-4_14

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  • DOI: https://doi.org/10.1007/978-3-642-56853-4_14

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-63214-3

  • Online ISBN: 978-3-642-56853-4

  • eBook Packages: Springer Book Archive

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