Abstract
Microchips that perform single cell capture, array, and identification have become powerful tools for single cell studies, which can reveal precise underlying mechanisms among bulk cell populations. However, current single cell capture and on-chip immunostaining methods consume more time and reagent than desired. To optimize this technology, we designed a novel trap structure for single cell capture, array, and release, and meanwhile an oscillatory method was used to perform rapid on-chip cell immunostaining. The trap structure array used equal distribution of lateral flow to achieve single cell array in high velocity flows and decrease the risk of clogging. A length of glass capillary with a sealed bubble was inserted into the outlet so that it could act in a manner analogous to that of a capacitor in an RC circuit. By applying one periodic air pressure to the inlet, oscillation motion was generated, which significantly enhanced the on-chip reaction efficiency. In addition, the oscillation performance could be easily regulated by changing the length of the capillary. The trapped cells could maintain their positions during oscillation; hence, they were able to be tracked in real time. Through our trap microchip, 12 μm microbeads were successfully trapped to form a microarray with a capture efficiency of ∼92.7% and 2 μm microbeads were filtered. With an optimized oscillation condition (Ppush = 0.03 MPa, f = 1 Hz, L = 3 cm), fast on-chip immunostaining was achieved with the advantages of less time (5 min) and reagent (2 μl) consumption. The effectiveness of this method was demonstrated through quantitative microbead and qualitative Caco-2 cell experiments. The device is simple, flexible, and efficient, which we believe provides a promising approach to single cell heterogeneity studies, drug screening, and clinical diagnosis.