A rigid microfluidic chip for high-throughput fluorescence-activated cell sorting

Fluorescence-activated cell sorting (FACS) holds great promise for the separation of single cells or cell populations according to specific light scattering and fluorescent characteristics. Here, we present a new perspective on microfluidic FACS (μFACS) with predictable geometry, which meets the requirements of high-throughput analysis and sorting. Instead of the widely applied elastic polydimethylsiloxane (PDMS), a rigid epoxy resin chip was rapidly fabricated and irreversibly encapsulated to eliminate channel deformation (tenfold reduction) and enhance performance while meeting high pressure (>600 kPa) and high flow rate application scenarios. Fluorescence discrimination and particle differentiation were additionally validated in a self-contained μFACS system using calibration microspheres and mammalian cells. The μFACS chip and system were integrally optimized to achieve a minimum interval (0.58 ms) with a mean flow rate of 1.5 m/s. Ultimately, event recording and automated sorting were accomplished in real time while achieving a sorting efficiency of 87% at cell throughput of 8,000 events/s. This rigid chip for high-throughput μFACS, which is independent of the physical properties of cells could pave the way for cell screening in plasma samples for personalized medicine.