Anisotropic colloidal crystal particles (CCPs) have showed their great potential in biotechnology and structural materials due to their anisotropic shapes and tunable optical property. However, their controllable generation is still a challenge. Here, a novel microfluidic approach is developed to generate anisotropic CCPs. The microfluidic device is composed of an injection capillary and a collection capillary with available size and shape. Based on the device, the anisotropic particles with non-close-packed colloidal crystal structures are achieved by photo-polymerizing droplet templates in a confined collection capillary with different shapes and sizes. Moreover, anisotropic close-packed CCPs can be made from non-close-packed CCPs through a thermal process. It is demonstrated that the anisotropic CCPs in different sizes, structural colors and shapes (rods, cuboids and disks) can be generated. These distinguishable features of resultant particles make them ideal barcodes for high-throughput bioassays. In order to prove it, DNA multiplex detection is carried out. The experimental results indicate that achieved particles have a great encoding capacity and are highly practical for multiplex coding bioassays. Therefore, we believe that the anisotropic CCPs would be highly promising barcodes in biomedical applications, including high-throughput bioassays and cell culture research where multiplexing is needed. (c) 2014 Elsevier Inc. All rights reserved.