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Studying Membrane Properties Using Fluorescence Lifetime Imaging Microscopy (FLIM)

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Fluorescent Methods to Study Biological Membranes

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 13))

Abstract

Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool to investigate the structure and composition of biological membranes. A wide variety of fluorescent probes suitable for FLIM experiments have been described. These compounds differ strongly in the details of their incorporation into membranes and in their responses toward changes in the membrane composition. In this chapter, we discuss and compare different classes of fluorescent membranes probes and their applications to studying biological membranes. We devote a section to a detailed description of fluorescent molecular rotors and their application to measuring local viscosity. As Förster resonance energy transfer (FRET) can be directly measured by changes in the donor fluorescence lifetime, FLIM is a very robust method to determine the distances between FRET pairs or the local concentrations of FRET-based membrane probes. Thus, we also discuss advantages and challenges of FRET-FLIM in the context of biological membranes. As biological membranes are considerably dynamic systems, imaging speed is often the limiting factor in biological FLIM experiments. Thus, novel fast imaging approaches and analysis methods to alleviate the issue of low photon statistics are also presented.

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Authors and Affiliations

  1. Nanobiophysics, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, 217, Enschede, 7500 AE, The Netherlands

    Martin T. Stöckl (Faculty of Science and Technology) & Vinod Subramaniam (Faculty of Science and Technology)

  2. Istituto di Biofisica, Consiglio Nazionale delle Ricerche (IBF-CNR), via Moruzzi 1, Pisa, 56124, Italy

    Ranieri Bizzarri

Authors
  1. Martin T. Stöckl
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  2. Ranieri Bizzarri
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  3. Vinod Subramaniam
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Correspondence to Martin T. Stöckl .

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Editors and Affiliations

  1. Faculté de Pharmacie, Laboratoire de Biophotonique, Université de Strasbourg, route du Rhin 74, Illkirch, 67401, France

    Yves Mély

  2. Faculté de Pharmacie, Laboratoire de Biophotonique et, Université de Strasbourg, route du Rhin 74, Illkirch, 67401, France

    Guy Duportail

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Stöckl, M.T., Bizzarri, R., Subramaniam, V. (2012). Studying Membrane Properties Using Fluorescence Lifetime Imaging Microscopy (FLIM). In: Mély, Y., Duportail, G. (eds) Fluorescent Methods to Study Biological Membranes. Springer Series on Fluorescence, vol 13. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2012_48

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