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Related Concept Videos

Enlargement of the Plasma Membrane01:22

Enlargement of the Plasma Membrane

Cell division and enlargement are processes that require precise control. The control ensures that cell division cannot proceed unless the cell has grown to a specific size. A spherical, dividing cell requires an approximately 1.6X increase in its surface area to double its volume. The secretory pathway also has a significant role in cell membrane enlargement. Secretory vesicles that bud off from the Golgi apparatus and later fuse with the plasma membrane during exocytosis are a major source of...
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The membrane comprises a group of distinct proteins responsible for carrying out a cell's specific function. For example, the plasma membrane of the human sperm, or a single germ cell, contains a unique set of proteins in the anterior...

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Related Experiment Video

Updated: May 22, 2026

Reconstitution of a Transmembrane Protein, the Voltage-gated Ion Channel, KvAP, into Giant Unilamellar Vesicles for Microscopy and Patch Clamp Studies
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Reconstitution of a Transmembrane Protein, the Voltage-gated Ion Channel, KvAP, into Giant Unilamellar Vesicles for Microscopy and Patch Clamp Studies

Published on: January 22, 2015

Elucidating membrane structure and protein behavior using giant plasma membrane vesicles.

Erdinc Sezgin1, Hermann-Josef Kaiser, Tobias Baumgart

  • 1Biophysics/BIOTEC, Technische Universität Dresden, Dresden, Germany.

Nature Protocols
|May 5, 2012
PubMed
Summary

Researchers can now observe phase separation in intact plasma membranes using giant plasma membrane vesicles (GPMVs). This breakthrough aids the study of membrane rafts and eukaryotic membrane lateral heterogeneity.

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Last Updated: May 22, 2026

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Area of Science:

  • Cell Biology
  • Biophysics
  • Membrane Biology

Background:

  • Eukaryotic plasma membranes exhibit lateral heterogeneity, with specialized domains known as membrane rafts playing crucial roles in cellular processes.
  • Understanding the dynamics and composition of membrane rafts is essential for deciphering cell signaling and membrane organization.
  • Giant plasma membrane vesicles (GPMVs) offer a model system to study membrane properties ex vivo.

Purpose of the Study:

  • To present a protocol for observing phase separation in intact plasma membranes using GPMVs.
  • To enable the study of membrane raft formation and lipid-protein interactions in a controlled environment.
  • To provide methods for quantitative analysis of membrane heterogeneity.

Main Methods:

  • Isolation of GPMVs from various cell types using chemical vesiculants.
  • Fluorescent labeling of vesicle components for microscopic visualization.
  • Temperature-controlled microscopy to induce and observe lipid phase separation below the miscibility transition temperature.
  • Analysis techniques including laurdan fluorescence spectroscopy, two-photon microscopy, and quantitative confocal microscopy.

Main Results:

  • Successful isolation and preparation of GPMVs from diverse cell types.
  • Demonstration of microscopic phase separation in GPMVs upon cooling.
  • Quantification of component partitioning between liquid-ordered (raft) and liquid-disordered (non-raft) phases.
  • Protocol completion within a 4-hour timeframe.

Conclusions:

  • The GPMV system provides a robust and accessible method for investigating eukaryotic membrane lateral heterogeneity.
  • This approach facilitates detailed studies of membrane raft formation and dynamics.
  • The described protocol enables advanced fluorescence-based analyses of membrane composition and organization.