Single-Cell Microarray Chip with Inverse-Tapered Wells to Maintain High Ratio of Cell Trapping.

Abstract

A single-cell microarray (SCM) influenced by gravitational force is expected to be one of the simple methods in various fields such as DNA analysis and antibody production. After trapping the cells in the SCM chip, it is necessary to remove the liquid from the SCM to wash away the un-trapped cells on the chip and treat the reagents for analysis. The flow generated during this liquid exchange causes the trapped cells to drop out of conventional vertical wells. In this study, we propose an inverse-tapered well to keep trapped cells from escaping from the SCM. The wells with tapered side walls have a reduced force of flow toward the opening, which prevents trapped cells from escaping. The proposed SCM chip was fabricated using 3D photolithography and polydimethylsiloxane molding techniques. In the trapping experiment using HeLa cells, the cell residual rate increased more than two-fold for the SCM chip with the inverse-tapered well with a taper angle of 30° compared to that for the conventional vertical SCM chip after multiple rounds of liquid exchanges. The proposed well structure increases the number of trapped cells and decreases the cell dropout rate to improve the efficiency of cellular analysis.