Single -studies have been gaining growing interests owing to the fact that whole-cell population studies cannot accurately detect cellular heterogeneity presents among the whole cell population, limiting our standing in coordinated cell behaviors. However, traditional methods for studying single cells rely on macro-scale tools that are either labor intensive or difficult to operate. Thus, the development of a high-throughput yet simple method for single cell culture experiments is of great importance. Here, we report a novel microfluidic device with a dual-well (DW) design concept for high-efficiency single-cell capture in large microwells (～77%), which provides ample space for cells to proliferate and differentiate. The ability of our DW device to simultaneously allow for high-efficiency single cell capture and culture was realized using sets of small microwells for cell capture, paired with large microwells for cell culture. By decoupling the capture and culture functionalities in our device, we were able to flexibly adjust the size of the large culture microwells without compromising single-cell capture efficiency. We also demonstrated single-cell acquiring efficiency of KT98 mouse neural stem cells, A549 and MDA-MB-435 cancer cells, and single-cell colony formation assays using our device, showing the diverse and flexible functionality of the DW device in cell culture applications.
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