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Published online by Cambridge University Press: 15 March 2011
The use of vesicles as amplifiers in biosensors is receiving increasing attention. Because vesicles may entrap thousands of reporter molecules, strong signal amplification can be obtained if a small number of analytes can simply release the entrapped reporters. Surface immobilization of vesicles with sensitivities for different analytes could then provide for simultaneous amplified detection of a number of analytes on a single chip. To achieve this goal, vesicles must first be stably adsorbed to a surface, without rupture. We have varied vesicle composition and charge (phosphatidylcholine, phosphatidylcholine-phosphatidic acid at 4.6 molar ratio) and solution ionic strength, to study the adsorption of fluorescent vesicles to glass, gold, and gold modified with chemisorbed acetylcysteine. Surfaces were characterized with angle-resolved X-ray photoelectron spectroscopy (ARXPS), and vesicle integrity and behavior was studied using entrapped and lipophilic fluorescent markers either together or in separate experiments. Diffusion coefficients (by photobleaching recovery) and vesicle fusion (by energy transfer) were monitored using confocal fluorescence microscopy. Finally, as a “proof of principle”, release of a self-quenching entrapped reporter dye (calcein) by the detergent Triton X-100 was followed in real time.