The ability to manipulate and selectively position cells into patterns or distinct microenvironments is an important component of many single cell experimental methods and biological engineering applications. Although a variety of particles and cell patterning methods have been demonstrated, most of them deal with the patterning of cell populations, and are either not suitable or difficult to implement for the patterning of single cells. Here, we describe a bottom-up strategy for the micropatterning of cells and cell-sized particles. We have configured a micromanipulator system, in which a pneumatic microinjector is coupled to a holding pipette capable of physically isolating single particles and cells from different types, and positioning them with high accuracy in a predefined position, with a resolution smaller than 10 µm. Complementary DNA sequences were used to stabilize and hold the patterns together. The system is accurate, flexible, and easy-to-use, and can be automated for larger-scale tasks. Importantly, it maintains the viability of live cells. We provide quantitative measurements of the process and offer a file format for such assemblies.
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