This study addresses the need for antibacterial medication that can overcome the current problems of antibiotics. It does so by suggesting two 1,8-naphthalimides (NI1 and NI2) containing a pyridinium nucleus become attached to the imide-nitrogen atom via a methylene spacer. Those fluorescent derivatives are covalently bonded to the surface of a chloroacetyl-chloride-modified cotton fabric. The iodometric method was used to study the generation of singlet oxygen (¹O₂) by irradiation of KI in the presence of monomeric 1,8-naphthalimides and the dyed textile material. Both compounds generated reactive singlet oxygen, and their activity was preserved even after they were deposited onto the cotton fabric. The antibacterial activity of NI1 and NI2 in solution and after their covalent bonding to the cotton fabric was investigated. In vitro tests were performed against the model gram-positive bacteria B. cereus and gram-negative P. aeruginosa bacteria in dark and under light iradiation. Compound NI2 showed higher antibacterial activity than compound NI1. The light irradiation enhanced the antimicrobial activity of the compounds, with a better effect achieved against B. cereu