Morphometric image analysis of giant vesicles

a new tool for quantitative thermodynamics studies of phase-separation in lipid membranes

Peter Rasmussen Husen, Laura Arriaga, Francisco Monroy, John Hjort Ipsen, Luis Bagatolli

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

We have developed a strategy to determine lengths and orientations of tie lines in the coexistence region of liquid-ordered and liquid-disordered phases of cholesterol containing ternary lipid mixtures. The method combines confocal-fluorescence-microscopy image stacks of giant unilamellar vesicles (GUVs), a dedicated 3D-image analysis, and a quantitative analysis based in equilibrium thermodynamic considerations. This approach was tested in GUVs composed of
1,2-dioleoyl-sn-glycero-3-phosphocholine/1,2-palmitoyl-sn-glycero-3-phosphocholine/cholesterol. In general, our results show a reasonable agreement with previously reported data obtained by other methods. For example, our computed tie lines were found to be nonhorizontal, indicating a difference in cholesterol content in the coexisting phases. This new, to our knowledge, analytical strategy offers a way to further exploit fluorescence-microscopy experiments in GUVs, particularly retrieving quantitative data for the construction of three lipid-component-phase diagrams containing cholesterol.
OriginalsprogEngelsk
TidsskriftBiophysical Journal
Vol/bind103
Udgave nummer11
Sider (fra-til)2304-2310
ISSN0006-3495
DOI
StatusUdgivet - 2012

Fingeraftryk

Membrane Lipids
Phase separation
Unilamellar Liposomes
Image analysis
Cholesterol
Thermodynamics
Fluorescence microscopy
Lipids
Phosphorylcholine
Confocal microscopy
Liquids
Phase diagrams
Chemical analysis
Experiments

Citer dette

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title = "Morphometric image analysis of giant vesicles: a new tool for quantitative thermodynamics studies of phase-separation in lipid membranes",
abstract = "We have developed a strategy to determine lengths and orientations of tie lines in the coexistence region of liquid-ordered and liquid-disordered phases of cholesterol containing ternary lipid mixtures. The method combines confocal-fluorescence-microscopy image stacks of giant unilamellar vesicles (GUVs), a dedicated 3D-image analysis, and a quantitative analysis based in equilibrium thermodynamic considerations. This approach was tested in GUVs composed of1,2-dioleoyl-sn-glycero-3-phosphocholine/1,2-palmitoyl-sn-glycero-3-phosphocholine/cholesterol. In general, our results show a reasonable agreement with previously reported data obtained by other methods. For example, our computed tie lines were found to be nonhorizontal, indicating a difference in cholesterol content in the coexisting phases. This new, to our knowledge, analytical strategy offers a way to further exploit fluorescence-microscopy experiments in GUVs, particularly retrieving quantitative data for the construction of three lipid-component-phase diagrams containing cholesterol.",
author = "Husen, {Peter Rasmussen} and Laura Arriaga and Francisco Monroy and Ipsen, {John Hjort} and Luis Bagatolli",
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Morphometric image analysis of giant vesicles : a new tool for quantitative thermodynamics studies of phase-separation in lipid membranes. / Husen, Peter Rasmussen; Arriaga, Laura; Monroy, Francisco; Ipsen, John Hjort; Bagatolli, Luis.

I: Biophysical Journal, Bind 103, Nr. 11, 2012, s. 2304-2310.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Morphometric image analysis of giant vesicles

T2 - a new tool for quantitative thermodynamics studies of phase-separation in lipid membranes

AU - Husen, Peter Rasmussen

AU - Arriaga, Laura

AU - Monroy, Francisco

AU - Ipsen, John Hjort

AU - Bagatolli, Luis

PY - 2012

Y1 - 2012

N2 - We have developed a strategy to determine lengths and orientations of tie lines in the coexistence region of liquid-ordered and liquid-disordered phases of cholesterol containing ternary lipid mixtures. The method combines confocal-fluorescence-microscopy image stacks of giant unilamellar vesicles (GUVs), a dedicated 3D-image analysis, and a quantitative analysis based in equilibrium thermodynamic considerations. This approach was tested in GUVs composed of1,2-dioleoyl-sn-glycero-3-phosphocholine/1,2-palmitoyl-sn-glycero-3-phosphocholine/cholesterol. In general, our results show a reasonable agreement with previously reported data obtained by other methods. For example, our computed tie lines were found to be nonhorizontal, indicating a difference in cholesterol content in the coexisting phases. This new, to our knowledge, analytical strategy offers a way to further exploit fluorescence-microscopy experiments in GUVs, particularly retrieving quantitative data for the construction of three lipid-component-phase diagrams containing cholesterol.

AB - We have developed a strategy to determine lengths and orientations of tie lines in the coexistence region of liquid-ordered and liquid-disordered phases of cholesterol containing ternary lipid mixtures. The method combines confocal-fluorescence-microscopy image stacks of giant unilamellar vesicles (GUVs), a dedicated 3D-image analysis, and a quantitative analysis based in equilibrium thermodynamic considerations. This approach was tested in GUVs composed of1,2-dioleoyl-sn-glycero-3-phosphocholine/1,2-palmitoyl-sn-glycero-3-phosphocholine/cholesterol. In general, our results show a reasonable agreement with previously reported data obtained by other methods. For example, our computed tie lines were found to be nonhorizontal, indicating a difference in cholesterol content in the coexisting phases. This new, to our knowledge, analytical strategy offers a way to further exploit fluorescence-microscopy experiments in GUVs, particularly retrieving quantitative data for the construction of three lipid-component-phase diagrams containing cholesterol.

U2 - 10.1016/j.bpj.2012.10.031

DO - 10.1016/j.bpj.2012.10.031

M3 - Journal article

VL - 103

SP - 2304

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JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

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