Making Giant Unilamellar Vesicles via Hydration of a Lipid Film

This unit describes protocols for making giant unilamellar vesicles (GUVs) based on rehydration of dried lipid films. These model membranes are useful for determining the impact of membrane and membrane‐binding components on lipid bilayer stiffness and phase behavior. Due to their large size, they are especially amenable to studies using fluorescence and light microscopy, and may also be manipulated for mechanical measurements with optical traps or micropipets. In addition to their use in encapsulation, GUVs have proven to be useful model systems for studying many cellular processes, including tubulation, budding, and fusion, as well as peptide insertion. The introduction of enzymes or proteins can result in reorganization, leading to such diverse behavior as vesicle aggregation, fusion, and fission. Curr. Protoc. Cell Biol. 40:24.3.1‐24.3.13. © 2008 by John Wiley & Sons, Inc.

[1]  L. Bagatolli,et al.  To see or not to see: lateral organization of biological membranes and fluorescence microscopy. , 2006, Biochimica et biophysica acta.

[2]  B. Stottrup,et al.  Sterol structure determines miscibility versus melting transitions in lipid vesicles. , 2005, Biophysical journal.

[3]  Daniel J. Estes,et al.  Electroformation of giant liposomes from spin-coated films of lipids. , 2005, Colloids and surfaces. B, Biointerfaces.

[4]  Petra Schwille,et al.  Sterol structure determines the separation of phases and the curvature of the liquid-ordered phase in model membranes. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[5]  M. Angelova,et al.  Phospholipase A2 promotes raft budding and fission from giant liposomes. , 2004, Chemistry and physics of lipids.

[6]  B. Nordén,et al.  Membrane binding and translocation of cell-penetrating peptides. , 2004, Biochemistry.

[7]  M. Dogterom,et al.  Membrane tube formation from giant vesicles by dynamic association of motor proteins , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Bruno Antonny,et al.  Lipid packing sensed by ArfGAP1 couples COPI coat disassembly to membrane bilayer curvature , 2003, Nature.

[9]  Sarah L Veatch,et al.  Separation of liquid phases in giant vesicles of ternary mixtures of phospholipids and cholesterol. , 2003, Biophysical journal.

[10]  Watt W. Webb,et al.  Imaging coexisting fluid domains in biomembrane models coupling curvature and line tension , 2003, Nature.

[11]  Sarah L Veatch,et al.  Organization in lipid membranes containing cholesterol. , 2002, Physical review letters.

[12]  P. Kinnunen,et al.  Observation of topical catalysis by sphingomyelinase coupled to microspheres. , 2002, Journal of the American Chemical Society.

[13]  E Gratton,et al.  Lipid rafts reconstituted in model membranes. , 2001, Biophysical journal.

[14]  P. Bassereau,et al.  A minimal system allowing tubulation with molecular motors pulling on giant liposomes , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[15]  P. Kinnunen,et al.  Vectorial budding of vesicles by asymmetrical enzymatic formation of ceramide in giant liposomes. , 2000, Biophysical journal.

[16]  J. Korlach,et al.  Characterization of lipid bilayer phases by confocal microscopy and fluorescence correlation spectroscopy. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[17]  E. Ikonen,et al.  Functional rafts in cell membranes , 1997, Nature.

[18]  R N Zare,et al.  Rapid preparation of giant unilamellar vesicles. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[19]  P. Mueller,et al.  Formation and properties of cell-size lipid bilayer vesicles. , 1983, Biophysical journal.

[20]  E Gratton,et al.  Two photon fluorescence microscopy of coexisting lipid domains in giant unilamellar vesicles of binary phospholipid mixtures. , 2000, Biophysical journal.

[21]  M. Angelova,et al.  Preparation of giant vesicles by external AC electric fields. Kinetics and applications , 1992 .