Figure 1.

Workflow and devices for droplet-based, single-cell, one-step RT-PCR. (a) Process flow of the platform. Cells suspended in an isotonic solution enter a microfluidic device through a channel separate from the RT-PCR reagents and lysis buffer. These channels fuse to a single orifice that injects into a channel met by opposing streams of oil to form monodispersed, 1 nL volume droplets. These droplets are collected in a PCR tube and thermal cycled to generate fluorescent amplification products. The droplets are then deposited in monolayers and analyzed using an automated imaging platform, after which the fluorescent signal in each droplet is quantified. (b) Droplet generator design (to scale) and brightfield image of the orifice of the device during droplet formation.