Use of air-liquid two-phase flow in hydrophobic microfluidic channels for disposable flow cytometers

This paper describes a disposable flow cytometer that uses an air-liquid two-phase microfluidic system to produce a focused high-speed liquid sample stream of particles and cells. The susceptibility of thin liquid columns to instabilities may suggest that focusing of sample liquids with streams of air would be difficult. The design of channel geometry, control of flow rates, and use of appropriate surface chemistries on the channel walls, however, enabled the generation of thin (15-100mum) and partially bounded sample streams that were stable and suitable for rapid cell analysis. Using an inverted epi-fluorescence microscope with a photo-multiplier tube, we demonstrated that the system is capable of counting the number of beads and C2C12 myoblast cells. The effects of different flow rates and surface chemistries of the channel walls on the air-liquid two-phase flows were characterized using optical and confocal microscopy. Use of air instead of liquids as a sheath fluid eliminates the need for large sheath liquid reservoirs, and reduces the volume and weight requirements. The low manufacturing cost and high volumetric efficiency make the air-sheath How cytometer attractive for use as a stand-alone device or as an integrated component of bio-artificial hybrid microsystems.