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Imaging membrane order using environmentally sensitive fluorophores.

G W Ashdown1, Dylan M Owen

  • 1Department of Physics, King's College London, Strand Campus, London, WC2R 2LS, UK.

Methods in Molecular Biology (Clifton, N.J.)
|October 22, 2014
PubMed
Summary
This summary is machine-generated.

Lipid membranes exist in ordered or disordered states, affecting protein interactions. Polarity-sensitive dyes like Laurdan can visualize these membrane phases in live cells using microscopy.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Membrane Biophysics

Background:

  • The lipid raft hypothesis posits that ordered and disordered lipid membranes regulate membrane-associated proteins.
  • Membrane order, quantified by the order parameter, influences lipid mobility and packing, impacting protein dynamics.
  • Disordered phases exhibit low-density packing, allowing water penetration, while ordered phases show condensation and water exclusion.

Purpose of the Study:

  • To explain how membrane order and disorder influence protein distribution and interactions.
  • To describe methods for quantifying membrane phase transitions.
  • To highlight the utility of polarity-sensitive fluorophores for live-cell imaging of membrane order.

Main Methods:

  • Utilizing polarity-sensitive fluorophores, such as Laurdan and di-4-ANEPPDHQ, embedded within lipid bilayers.
  • Employing various microscopy techniques to visualize and quantify membrane order distributions.
  • Measuring changes in fluorophore emission properties correlated with alterations in membrane phase.

Main Results:

  • Demonstrated that membrane phase transitions (ordered vs. disordered) are associated with distinct lipid packing densities and water penetration.
  • Showcased the ability of Laurdan and di-4-ANEPPDHQ to report on local membrane polarity and order.
  • Confirmed the applicability of these fluorescent probes for imaging membrane order heterogeneity in live cells.

Conclusions:

  • Membrane order is a critical biophysical parameter influencing cellular processes.
  • Polarity-sensitive fluorophores provide a powerful tool for studying dynamic changes in membrane organization.
  • Microscopy-based imaging with probes like Laurdan enables detailed analysis of membrane phase behavior in living systems.