Fluorescent liposome flow markers for microscale particle image velocimetry

Unilamellar liposomes carrying both encapsulated and surface-immobilized fluorophores have been synthesized as novel fluorescent markers to image flow profiles in microfabricated structures. The unilamellar liposomes were made with phospholipids and cholesterol by extrusion through a polycarbonate membrane, They contained carboxyfluorescein in the aqueous core and fluorescein-labeled lipids in the bilayer to render them both a surface and volume fluorescer, maximizing their fluorescence intensity. The lipid composition was chosen to impart a net negative charge to liposomes to minimize self-aggregation as well as interaction with negatively charged glass surfaces of the channels. These liposomes were monodisperse (mean diameter 283 nm), neutrally buoyant, and hydrophilic and exhibited no adsorption on glass surfaces. Unlike polystyrene spheres, they were readily broken up by surfactants, thereby allowing for easy and complete removal from microfluidic channels. The fluorescent liposomes were used to investigate pressure-driven now in an offset cross intersection in a microfluidic chip and provided images with excellent signal-to-noise ratio. A novel computational scheme that is particularly suitable for analyzing particle-image velocimetry data in micrometer-scale flow channels was employed to analyze the images. These liposomes are easily synthesized and can be custom-made for various applications to offer a broad range of surface and volume characteristics such as charge, size, and surface chemistry.