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Imaging orientational order and lipid density in multilamellar vesicles with multiplex CARS microscopy
Author(s) -
WURPEL G. W. H.,
RINIA H. A.,
MÜLLER M.
Publication year - 2005
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2005.01462.x
Subject(s) - vesicle , microscopy , chemistry , lipid bilayer , multiplex , polarization microscopy , raman scattering , phosphatidylcholine , raman spectroscopy , analytical chemistry (journal) , biophysics , chemical physics , phospholipid , optics , membrane , chromatography , physics , biochemistry , biology , bioinformatics
Summary Multiplex coherent anti‐Stokes Raman scattering (CARS) microscopy is used to measure the width of the orientational order distribution of lipid acyl chains within a three‐dimensionally confined microscopic probing volume. A theoretical model is developed to describe and simulate the polarization‐dependent measurements. We observe that the orientational order in phosphatidylcholine multilammellar vesicles increases significantly upon addition of small amounts (≤ 15 mol%) of cholesterol and is significantly reduced for unsaturated lipids. Based on these measurements and using the quantitative nature of multiplex CARS microscopy the exact local concentration of lipid molecules within the vesicles can be measured in terms of the number of lipid bilayers present in the microscopic probing volume.