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Spectral Decomposition of Intracellular Complex Fluorescent Signals Using Multiwavelength Phase Modulation Lifetime Determination

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Abstract

Lifetime-based spectral decomposition using a frequency-domain phase/modulation technique has been developed on our UV confocal microspectrofluorimeter prototype. Examples of such an application are first given in the case of complex mixtures of three overlapping fluorescent components in solution. Preliminary results concerning cellular potential application are also reported in the case of Snarf-1 pH sensor. Limits and perspectives of such an approach at the subcellular level are discussed in view of future improvement.

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Authors and Affiliations

  1. Institute of Physics, Charles University, Ke Karlovu 5, 12116, Prague 2, Czech Republic

    P. Praus

  2. Laboratoire de Physicochimie Biomoléculaire et Cellulaire (CNRS ESA 7033), Université P. et M. Curie, Case 138, 4 Place Jussieu, F-75252, Paris Cedex 05, France

    F. Sureau

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  1. P. Praus
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  2. F. Sureau
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Praus, P., Sureau, F. Spectral Decomposition of Intracellular Complex Fluorescent Signals Using Multiwavelength Phase Modulation Lifetime Determination. Journal of Fluorescence 10, 361–364 (2000). https://doi.org/10.1023/A:1009426429458

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