Protein nanoarrays for high-resolution patterning of bacteria on gold surfaces

In recent years, the majority of research on surface patterning, as a means of precisely controlling cell -positioning and adhesion on surfaces, has focused on eukaryotic cells. Such research has led to new insights into cell biology, advances in tissue engineering, and cell motility. In contrast, considerably less work has been reported on tightly controlled patterning of bacteria, despite its potential in a wide variety of applications, including fabrication of in vitro model systems for studies of bacterial processes, such as quorum sensing and horizontal gene transfer. This is partly due to their small size - often 1-3 μm or less. To study these processes, microscale and nanoscale engineered material surfaces must be developed to create in vitro bacteria arrays, which can allow valuable insights into natural systems such as the soil or the human gut, and are often complex and spatially structured habitats. Here, we outline a protocol to create defined patterns of bacteria to study such systems at the single cell level that is based on the formation of protein nanoarrays on mannoside-terminated self-assembled monolayers via nanocontact printing and the subsequent deposition of bacteria from solution on the unpatterned regions of the mannoside-terminated substrate.