Optical pH Monitoring in Microdroplet Platforms for Live Cell Experiments Using Colloidal Surfactants

Droplet microfluidic technology facilitates the development of high-throughput screening applications in nanoliter volumes. Surfactants provide stability for emulsified monodisperse droplets to carry out compartmentalization. Fluorinated silica-based nanoparticles are used; they can minimize crosstalk in microdroplets and provide further functionalities by surface labeling. Here we describe a protocol for monitoring pH changes in live single cells by fluorinated silica nanoparticles, for their synthesis, chip fabrication, and optical monitoring on the microscale. The nanoparticles are doped with ruthenium-tris-1,10-phenanthroline dichloride on the inside and conjugated with fluorescein isothiocyanate on the surface. This protocol may be used more generally to detect pH changes in microdroplets. The fluorinated silica nanoparticles can also be used as droplet stabilizers with an integrated luminescent sensor for other applications.