Customizable multi-well plates for high-throughput 2D and 3D laser direct-write

Laser direct-write (LDW) is a forward transfer deposition technique that has recently been adapted for 2D/3D printing of biological materials. LDW is built upon a CAD/CAM platform and scalable technologies, making it an attractive biofabrication tool for many applications, such as high-throughput in vitro testing. To truly realize these capabilities, LDW must be able to print into multi-well plates, (e.g., 24-, 48-, 96-well plates), for analysis of multiple samples in parallel (e.g., spectrophotometry, microscopy, RT-PCR), as such applications demand for high throughput. However, the geometries of standard multi-well plates interfere with the LDW print ribbon, requiring a much greater transfer distance (> 10mm between ribbon and receiving substrate). This large ribbon-substrate distance compromises print quality significantly (e.g., loss of print registry, generation of satellite beads, scattered print spots/droplets). Moreover, the wells cannot be spin-coated evenly, leading to non-uniform substrates. To overcome these hurdles, we developed a novel method to produce customizable multi-well substrates that can be spin-coated, and do not restrict ribbon-substrate spacing. This technique can be used to create LDW-compatible substrates with custom multi-well geometries, to greatly improve the flexibility, utility, and throughput of LDW.