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STED-FCS Nanoscopy of Membrane Dynamics

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Fluorescent Methods to Study Biological Membranes

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

Abstract

Lipid-lipid and lipid-protein interactions such as the formation of lipid nanodomains (often denoted “rafts”) are considered to play a functional part in a whole range of membrane-associated processes. However, their direct and noninvasive observation in living cells is impeded by the resolution limit of >200 nm of a conventional far-field optical microscope. With the superior spatial resolution of STED nanoscopy, it is now possible to directly resolve nanoscale membrane organization. While direct imaging of membrane heterogeneities turns challenging due to their fast dynamics, the combination of STED nanoscopy with tools such as fluorescence correlation spectroscopy (FCS) allows the disclosure of complex nanoscopic dynamical processes. By performing FCS measurements in observation spots tuned to a diameter of down to 30 nm, new details of molecular membrane dynamics have been obtained: Unlike fluorescent phosphoglycerolipids, fluorescent sphingolipids are transiently (~10 ms) trapped on the nanoscale in often cholesterol- and cytoskeleton-assisted molecular complexes. These interactions are distinct for different lipids and may play an important role in cellular functionality. Comparison of the trapping characteristics to the organization of the different fluorescent lipid analogs in model membranes reveals details of the role of lipid “rafts”. This chapter reviews how STED-FCS may shed new light on the role of lipid-protein interactions and nanodomains for membrane bioactivity.

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Acknowledgement

This work could not have been realized without the work of the people from the department of NanoBiophotonics: Christian Ringemann, Veronika Mueller, Alf Honigmann, Rebecca Medda, Vladimir Belov, Svetlana Polyakova, Birka Llaken, Class von Middendorf und Andreas Schönle, especially by the extraordinary support and interaction with Stefan Hell, as well as by the lipid syntheses and discussions with Günter Schwarzmann (University Bonn). Further fruitful discussions with Herve Rigneault (Marseille), the Simons group (MPI Dresden), and the Schwille group (Bioquant Dresden) are greatly acknowledged.

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  1. Department of NanoBiophotonics, Max-Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen, 37077, Germany

    Christian Eggeling

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  1. Christian Eggeling
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Correspondence to Christian Eggeling .

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  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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Eggeling, C. (2012). STED-FCS Nanoscopy of Membrane Dynamics. 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_50

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