Abstract
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.
Highlights
The manipulation and patterning of individual micro-objects is a critical technology for micro-assembly [1,2] and many biomedical applications [3,4]
Our method offers an advantage over other patterning techniques, as it combines single cell patterning and precise positioning with the ability to selectively choose the particles that are positioned, while keeping the cells viable
Holding micropipettes were connected to the microinjector and used to grab and dispense single cells (Figure 1B,C)
Summary
The manipulation and patterning of individual micro-objects is a critical technology for micro-assembly [1,2] and many biomedical applications [3,4]. An important goal in tissue engineering and the study of the replication of tissue micro-environments is the development of a technique to position different types of cells and extracellular elements precisely into a specific predefined position with a corresponding resolution, keeping the cells viable and functioning [5,6]. Such a method will make it possible to study cell biology at a single cell level, by examining single cell interactions with drugs, growth factors, or external stimuli. The principle of the bottom-up approach is to pattern individual components of a tissue according to a predefined pattern, which will guide the maturation of cell self-sorting and self-assembly capabilities, as well as the morphogenesis mechanism [16]
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