User-friendly universal and durable subcellular-scaled template for protein binding: Application to single-cell patterning

A new method for subcellular-sized protein patterning on a SiOx substrate is demonstrated by dip-pen nanolithography printed aldehyde-terminated alkylsilane template. The aldehyde-silane template is stable and durable; for example, subcellular scaled IgG protein array can be obtained using one-year old aldehyde-silane template. Moreover, single cell patterning is successfully carried out by extracellular material (ECM) protein microarray and nanoarray fabricated on an aldehyde-silane template. With more than half of chance, single- or double-cells are successfully attached on fibronectin protein nanoarrays in 21 × 21 μm ² (7 × 7 dot array) and 42 × 42 μm² (14 × 14 dot array). The fibronectin nanoarray with small area (21 × 21 μm²) shows the more rate of single cell attachment. Therefore, it is also demonstrated that cell patterning can be controlled by adjusting the nanostructure of ECM materials.