High throughput single cell separation and identification using a self-priming isometric and Equant screw valve-based (SIES) microfluidic chip.

The emergence of various single cell separation and identification platforms has greatly promoted the development of single cell research. Among these platforms, microfluidic chip-based strategies occupy a significant position in single cell separation and identification. Here, we proposed a self-priming isometric and Equant screw valve-based microfluidic chip (SIES chip) for high throughput single cell isolation and identification. With several special designs, such as a peripheral water tank to balance negative pressure distribution in a marginal area of the chip, a screw valve to preserve the suction power during the step-by-step sample loading, and multistage branching "T" shape channels to separate cells evenly into the chambers, up to 2000 single cells can be well dispersed and analyzed at the same time using this chip. We applied this chip for the isolation and identification of single A549 cells targeting the activated leukocyte cell adhesion molecule (ALCAM) gene. The results showed that only a small proportion (approximately 5.1%) of A549 cells expressed ALCAM, which can potentially provide a reference for A549 cell reclassification. Besides being inexpensive, user-friendly and portable, our chip can be used in some resource-limited settings and may have a great potential in POC (Point-of-Care) applications.