Due to their antibacterial activity, chitosan‑carbon dot composites possess great potential for pharmaceuticals, medicine, and food preservation. Conducting a comprehensive study of the interactions between chitosan, carbon dots, and bacteria is crucial to understanding the processes behind applying these composites. This study aimed to immobilize carbon dots (C-dots) synthesized from Elaeagnus angustifolia fruits on chitosan and glass microbeads' surfaces, to characterize the test materials obtained after synthesis and immobilization, and to investigate their antibacterial potentials. C-dot synthesis was carried out from water extract in an acidic medium with the help of microwave irradiation, and their structural and optical properties were characterized by TEM, XRD, FT-IR, UV–vis, Zeta potential, and fluorescence methods. The surface of the glass microbeads was first activated and functionalized with surface amine groups with a silaning agent. C-dots were immobilized on both glass and chitosan microbeads using a crosslinking agent. Antibacterial potentials of nine different test materials, obtained before or after immobilization, were evaluated both qualitatively (MIC and MBC) and quantitatively (GI50) on E. coli, S. typhimurium, B. subtilis, and S. aureus, with the standard broth microdilution method. FT-IR and SEM-EDX analyses showed that C-dots were immobilized on chitosan (˂1 mm) and glass (˂100 μm) microbead surfaces. C-dots reduced the cell viability by ~25 % on S. typhimurium and B. subtilis (MIC = 25 mg/mL). It was also found that the highest antibacterial effect was recorded for C-dots-glass microbeads, which had a toxic effect of 43 % on S. aureus. In addition, binding C-dots to glass microbeads increased the antibacterial effect selectively in Gram-positive bacteria, while binding to chitosan microbeads was effective in all bacteria. The study showed that the antibacterial potential of C-dots-chitosan microbeads is more effective than C-dots-glass microbeads. C-dots could be used as carbon-based nanomaterials in antibacterial surface preparation once immobilized.