A simple microwell-device for isolation and analysis of single cell

Abstract

Circulating tumor cells (CTCs) are cells that have shed into the vasculature from a primary tumor and circulate in the bloodstream. Since individual CTCs are heterogeneous cells which have a variety of biological and physical properties such as EpCAM expression and different size, single analysis of CTCs has emerged as promising diagnostic and prognostic tool for patients with metastatic cancers. This paper describes a microfluidic-based droplet generation method, for encapsulating a single circulating tumor cells(CTCs) into a picoliter- or femtoliter-volume aqueous droplet that is surrounded by an immiscible phase. Once PBS with the target cells is injected into an inlet with a conventional micropipette. Injected cells are settled on bottom of channel because of gravity, for several minutes. Then, the exterior with cells of micro-well removed by the injection of oil, the rest of them trapped in micro well were encapsulated as well. We successfully trap a single cell in the well and generate a droplet by injecting oil. A droplet formed in each well can be used to trap and analyze a single CTCs. We also found out that the best trapping efficiency of single cells was 74% at 21um well.