Preparing giant unilamellar vesicles (GUVs) of complex lipid mixtures on demand: Mixing small unilamellar vesicles of compositionally heterogeneous mixtures
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Preparing giant unilamellar vesicles (GUVs) of complex lipid mixtures on demand : Mixing small unilamellar vesicles of compositionally heterogeneous mixtures. / Bhatia, Tripta; Husen, Peter; Brewer, Jonathan; Bagatolli, Luis A.; Hansen, Per L.; Ipsen, John H.; Mouritsen, Ole G.
In: Biochimica et Biophysica Acta - Biomembranes, Vol. 1848, No. 12, 2015, p. 3175-3180.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Preparing giant unilamellar vesicles (GUVs) of complex lipid mixtures on demand
T2 - Mixing small unilamellar vesicles of compositionally heterogeneous mixtures
AU - Bhatia, Tripta
AU - Husen, Peter
AU - Brewer, Jonathan
AU - Bagatolli, Luis A.
AU - Hansen, Per L.
AU - Ipsen, John H.
AU - Mouritsen, Ole G.
PY - 2015
Y1 - 2015
N2 - Giant unilamellar vesicles (GUVs) are simple model membrane systems of cell-size, which are instrumental to study the function of more complex biological membranes involving heterogeneities in lipid composition, shape, mechanical properties, and chemical properties. We have devised a method that makes it possible to prepare a uniform sample of ternary GUVs of a prescribed composition and heterogeneity by mixing different populations of small unilamellar vesicles (SUVs). The validity of the protocol has been demonstrated by applying it to ternary lipid mixture of DOPC, DPPC, and cholesterol by mixing small unilamellar vesicles (SUVs) of two different populations and with different lipid compositions. The compositional homogeneity among GUVs resulting from SUV mixing is quantified by measuring the area fraction of the liquid ordered-liquid disordered phases in giant vesicles and is found to be comparable to that in GUVs of the prescribed composition produced from hydration of dried lipids mixed in organic solvent. Our method opens up the possibility to quickly increase and manipulate the complexity of GUV membranes in a controlled manner at physiological buffer and temperature conditions. The new protocol will permit quantitative biophysical studies of a whole new class of well-defined model membrane systems of a complexity that resembles biological membranes with rafts.
AB - Giant unilamellar vesicles (GUVs) are simple model membrane systems of cell-size, which are instrumental to study the function of more complex biological membranes involving heterogeneities in lipid composition, shape, mechanical properties, and chemical properties. We have devised a method that makes it possible to prepare a uniform sample of ternary GUVs of a prescribed composition and heterogeneity by mixing different populations of small unilamellar vesicles (SUVs). The validity of the protocol has been demonstrated by applying it to ternary lipid mixture of DOPC, DPPC, and cholesterol by mixing small unilamellar vesicles (SUVs) of two different populations and with different lipid compositions. The compositional homogeneity among GUVs resulting from SUV mixing is quantified by measuring the area fraction of the liquid ordered-liquid disordered phases in giant vesicles and is found to be comparable to that in GUVs of the prescribed composition produced from hydration of dried lipids mixed in organic solvent. Our method opens up the possibility to quickly increase and manipulate the complexity of GUV membranes in a controlled manner at physiological buffer and temperature conditions. The new protocol will permit quantitative biophysical studies of a whole new class of well-defined model membrane systems of a complexity that resembles biological membranes with rafts.
KW - Confocal fluorescence microscopy
KW - Domains
KW - Giant unilamellar vesicles
KW - Lipid heterogeneity
KW - Membranes
KW - Rafts
U2 - 10.1016/j.bbamem.2015.09.020
DO - 10.1016/j.bbamem.2015.09.020
M3 - Journal article
C2 - 26417657
AN - SCOPUS:84944206072
VL - 1848
SP - 3175
EP - 3180
JO - B B A - Biomembranes
JF - B B A - Biomembranes
SN - 0005-2736
IS - 12
ER -
ID: 230974315