There is a growing interest on the fabrication of bacteria and virus microarray owing to their great potential in many biological applications ranging from diagnostic devices to advanced platforms for fundamental studies on molecular biology. Over the past decade, a number of studies with regard to the biomolecular patterning have been presented. Capillary force lithography (CFL) for polymeric thin films can provide well-ordered microarray structures over a large area in a facile and cost-efficient way while maintaining its biocompatibility during a process. Patterned polymeric structures can be utilized either to physical barriers for the confinement of bacteria or to physicochemical template for the subsequent binding of viruses. In this chapter, we have shown that the patterned structures of poly(ethylene glycol) (PEG) containing polymer enables a selective binding of Escherichia coli , leading to a physically guided microarray of bacteria. Additionally, we demonstrate the fabrication of virus microarray of M13 viruses via electrostatic interactions with a prepatterned microstructure of polyelectrolyte multilayers.