Drop-on-demand inkjet-based cell printing with 30-μm nozzle diameter for cell-level accuracy

We present drop-on-demand inkjet-based mammalian cell printing with a 30-μm nozzle diameter for cell-level accuracy. High-speed imaging techniques have been used to analyze the go-and-stop movement of cells inside the nozzle under a pulsed pressure generated by a piezo-actuator and the jet formation after ejection. Patterning of an array of 20 × 20 dots on a glass substrate reveals that each printed drop contains 1.30 cells on average at the cell concentration of 5.0 × 10⁶ cells ml^-1 for the very small nozzle, whereas larger nozzles with the diameter of 50 and 80 μm deliver 2.57 and 2.88 cells per drop, respectively. The effects of the size and concentration of printed cells on the number of cells have also been investigated. Furthermore, the effect of the nozzle diameter on printed cells has been evaluated through an examination of viability, proliferation, and morphology of cells by using a live/dead assay kit, CCK-8 assay, and cellular morphology imaging, respectively. We believe that the 30-μm inkjet nozzle can be used for precise cell deposition without any damages to the printed mammalian cells.