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Atomic Force Microscopy Imaging and Force Spectroscopy of Supported Lipid Bilayers
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In situ imaging of detergent-resistant membranes by atomic force microscopy.

M C Giocondi1, V Vié, E Lesniewska

  • 1Centre de Biochimie Structurale, INSERM U414, 29 rue de Navacelles, Montpellier Cedex, France.

Journal of Structural Biology
|August 18, 2000
PubMed
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Detergent-resistant membranes (DRMs) studied in cells appear larger than native domains, suggesting rearrangement during Triton X-100 treatment. These in situ images reveal DRMs associate with the cytoskeleton.

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Area of Science:

  • Cell Biology
  • Biophysics

Background:

  • Detergent-resistant membranes (DRMs) are crucial for studying plasma membrane domains.
  • The in situ size and native correspondence of DRMs are debated.
  • Classical microscopy cannot determine the in situ size of DRMs.

Purpose of the Study:

  • To determine the in situ size of detergent-resistant membrane domains.
  • To investigate the relationship between native membrane microdomains and isolated DRMs.
  • To explore the association of DRMs with the cytoskeleton in situ.

Main Methods:

  • Tapping mode atomic force microscopy (AFM) was used to obtain in situ 3D images.
  • Triton X-100-treated cells were imaged in buffer.
  • AFM allowed visualization of fragile cellular structures.

Main Results:

  • In situ images revealed detergent-treated plasma membrane fragments forming domains larger than 15-20 microm(2).
  • This observed size is significantly larger than previously estimated native microdomain sizes.
  • The cytoskeleton was visualized and appeared resistant to detergent extraction, associated with DRMs.

Conclusions:

  • Detergent-resistant membrane (DRM) domains likely rearrange and enlarge during Triton X-100 treatment.
  • In situ DRMs are associated with the membrane cytoskeleton.
  • AFM provides a novel method for studying membrane structures in their native-like state.