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Imaging orientational order and lipid density in multilamellar vesicles with multiplex CARS microscopy
G W H Wurpel1, H A Rinia, M Müller
1Swammerdam Institute for Life Sciences, University of Amsterdam, PO Box 96062, 1090 GB Amsterdam, The Netherlands.
Journal of Microscopy
|April 9, 2005
Summary
Multiplex coherent anti-Stokes Raman scattering (CARS) microscopy reveals how cholesterol affects lipid order in vesicles. This technique quantifies lipid concentration by measuring bilayer presence within the probed volume.
Area of Science:
- Biophysics
- Chemical Physics
- Materials Science
Background:
- Understanding lipid acyl chain order is crucial for cell membrane function.
- Cholesterol and lipid unsaturation are known modulators of membrane properties.
Purpose of the Study:
- To measure the orientational order distribution of lipid acyl chains in confined volumes.
- To investigate the effect of cholesterol and unsaturated lipids on this order.
- To quantify local lipid concentration using multiplex CARS microscopy.
Main Methods:
- Utilized multiplex coherent anti-Stokes Raman scattering (CARS) microscopy.
- Developed a theoretical model for polarization-dependent measurements.
- Applied the technique to phosphatidylcholine multilammellar vesicles.
Main Results:
- Observed increased orientational order with cholesterol addition (
- Found reduced orientational order with unsaturated lipids.
- Demonstrated quantitative measurement of lipid concentration based on bilayer number.
Conclusions:
- Multiplex CARS microscopy provides quantitative insights into lipid acyl chain order.
- Cholesterol significantly enhances lipid order in phosphatidylcholine vesicles.
- The method allows for precise determination of local lipid concentration in membrane systems.