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Quantitative Composition Analysis of Lipid Membranes by High-Resolution Secondary Ion Mass Spectrometry

P. K. WeberW. C. LinI. D. HutcheonM. L. KraftM. L. LongoS. G. BoxerC. D. Blanchette

出版	United States. Department of Energy, 2005
URL	http://books.google.com.hk/books?id=iRVunQAACAAJ&hl=&source=gbs_api

註釋The lateral organization and interactions of lipid and protein components within biological membranes are essential for their functions. Investigations of the lateral organization within membranes hinge upon the ability to differentiate one component of interest from another. Typically, fluorophores are conjugated to specific components, and the organization is probed with fluorescence microscopy. However, bulky fluorophores may change the physical properties of the components they label, only the labeled components can be visualized, and the diffraction limit of light restricts the lateral resolution. Here we present a method to image microdomains within supported lipid membranes using isotopic labels and high-resolution secondary ion mass spectrometry (SIMS) performed with the NanoSIMS 50 (Cameca). Lateral resolution of 100 nm is achieved with high sensitivity. Quantitative information on the lipid composition within each domain was determined using calibration curves constructed from homogeneous lipid bilayer samples that systematically varied in the isotopically labeled lipid content.