Size and fluorescence measurements of individual droplets by flow cytometry

Abstract

Biotechnological applications of emulsions, such as micro-reactors or drug carriers, demand accurate characterization techniques, able to measure the size and biochemical content of the droplets at the individual level. Since no available characterization technique completely fulfills these needs, we extended the use of flow cytometry, which was originally developed for cell studies, to the straightforward and quantitative characterization of micron-sized emulsions. Our method determines the size of soybean oil droplets from flow cytometric measurements of forward scattering and side scattering intensities combined with the theoretical scattered intensities exactly derived from Mie theory and numerically integrated with respect to the optical setup of the instrument. We evaluate the accuracy of our method by comparing the size distribution obtained for a monodisperse emulsion sample to the corresponding distribution measured with a commercial instrument. Applied to emulsion droplets functionalized with fluorescent streptavidin, our method allows for monitoring of the rate of grafted molecules on interfaces with a precision never obtained before.