Development of novel antibacterial fibers with mass production is urgently required in the technical textiles industry. In this paper, a series of segmented-pie composite microfibers based on polyamide 6 (PA6) and different amounts of copper–carbon nanoparticles (CuCNPs) were fabricated by utilizing a melt-spinning apparatus with twin-screw extruders. The encapsulation of CuCNPs and the formation of segmented-pie structure of as-prepared PA6/CuCNP microfibers were confirmed. The CuCNPs or their agglomeration with an average diameter of approximately 200 nm exhibited a uniform distribution in PA6/CuCNP segmented-pie microfibers. Compared with the pure PA6 microfibers, the PA6/CuCNP segmented-pie microfibers showed obviously enhanced crystallinity, thermal stability as well as UV resistance. As the CuCNP content increased to 1.0 wt%, the tensile strength and initial modulus increased to 3.79 cN/dtex and 22.4 cN/dtex, respectively. Importantly, the PA6/CuCNP segmented-pie microfibers presented excellent antimicrobial activities to both Escherichia coli and Staphylococcus aureus (antimicrobial efficiency around 99%) and great antifungal activity to Candida albicans (antimicrobial efficiency around 82%). Taken together, our present study demonstrated that the PA6/CuCNP segmented-pie microfibers show great prospects in the fabrication of technical textiles for healthcare applications.