A quantitative determination of lipid bilayer deposition efficiency using AFM

利用原子力显微镜定量测定脂质双层沉积效率

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Abstract

The efficacy of a number of different methods for depositing a dimyristoylphosphatidylcholine (DMPC) lipid bilayer or DMPC-cholesterol (3 : 1) mixed bilayer onto a silicon substrate has been investigated in a quantitative manner using atomic force microscopy (AFM) image analysis to extract surface coverage. Complementary AFM-IR measurements were used to confirm the presence of the lipids. For the Langmuir-Blodgett/Schaefer deposition method at temperatures below the chain-melting transition temperature (T (m)), a large number of bilayer defects resulted when DMPC was deposited from a water subphase. Addition of calcium ions to the trough led to smaller, more frequent defects, whereas addition of cholesterol to the lipid mixture led to a vast improvement in bilayer coverage. Poor coverage was achieved for deposition at temperatures above T (m). Formation of the deposited bilayer from vesicle fusion proved a more reliable method for all systems, with formation of near-complete bilayers within 60 seconds at temperatures above T (m), although this method led to a higher probability of multilayer formation and rougher bilayer surfaces.

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