1D metallic nanostructures have become a subject of intensive research due to their potential application in the biomedical field such as sensing, imaging, drug delivery, and biodevices. Herein, anodic aluminum oxide (AAO) template is used to electrochemically grow the highly crystalline Cu nanowires. The electrodeposition potential for copper solution is identified by cyclic voltammetry. The Cu nanowire‐deposited AAO template is further characterized using field emission scanning electron microscope (FESEM), energy dispersive X‐ray (EDX) spectroscopy, wavelength dispersive X‐ray fluorescence (WDXRF) spectroscopy, X‐ray diffraction (XRD), and electrochemical impedance spectroscopy (EIS). The Cu‐deposited AAO template is tested for antibacterial activity against different microorganisms (including both gram positive as well as gram negative). Further, the assessment of the cytotoxic nature of Cu nanowires is a fundamental step for their application in the biomedical field. The favorable properties of Cu‐deposited AAO template in terms of excellent antibacterial effects and low cytotoxicity promote their application in the biomedical field, especially for medical diagnosis.