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Related Experiment Videos

Laser-assisted fluorescence microscopy for measuring cell membrane dynamics.

Herbert Schneckenburger1, Michael Wagner, Martina Kretzschmar

  • 1Fachhochschule Aalen, Institut fur Angewandte Forschung, Beethovenstr. 1, 73430 Aalen, Germany.

Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology
|August 6, 2004
PubMed
Summary

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This study uses advanced fluorescence microscopy to measure cell membrane stiffness and fluidity. Key parameters like generalized polarization and fluorescence lifetime reveal changes during cell growth and aging.

Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy Techniques

Background:

  • Cell membranes are dynamic structures whose physical properties influence cellular functions.
  • Understanding membrane stiffness and fluidity is crucial for cell physiology.
  • Advanced microscopy offers powerful tools to probe membrane characteristics at the molecular level.

Purpose of the Study:

  • To characterize living cell membranes using a combination of laser-assisted fluorescence microscopy techniques.
  • To establish generalized polarization (GP) and effective fluorescence lifetime (tau(eff)) as quantitative parameters for membrane stiffness and fluidity.
  • To investigate the influence of temperature, cell growth, and aging on membrane physical properties.

Main Methods:

  • Utilized microspectrofluorometry, total internal reflection fluorescence microscopy (TIRFM), fluorescence lifetime imaging (FLIM), and Förster resonance energy transfer (FRET) spectroscopy.

Related Experiment Videos

  • Employed the membrane marker laurdan to assess spectral shifts (GP) and fluorescence lifetimes (tau(eff)).
  • Investigated laurdan's proximity to a molecular acceptor (DiI) using non-radiative energy transfer.

    • Main Results:

    • Generalized polarization (GP) correlated with membrane stiffness, decreasing with temperature but increasing during cell growth.
    • Plasma membranes exhibited higher GP than intracellular membranes.
    • Distinct microdomains with varying fluorescence lifetimes (tau(eff)) were observed above 30°C, disappearing with cell aging.

    Conclusions:

    • GP and tau(eff) derived from laurdan fluorescence are effective parameters for quantifying cell membrane stiffness and fluidity.
    • Temperature, cell growth, and aging significantly impact membrane physical properties.
    • FRET-based measurements offer potential for probing membrane dynamics within specific microenvironments.