The study reports designing of a new, low-cost and environmentally friendly colorimetric and fluorometric sensor by using cellulose-based materials for detection and determination of Fe(III). To make powder cellulose (Cel) and filter paper (PCel) fluorescent, they were modified with hexamethylene diisocyanate (HMDI) and 4-sulfo-1,8-naphthalimide (Nap). Fluorescent Cel-Nap and PCel-Nap materials were used for spectroscopic detection of Fe(III). The working range of the designed sensor was determined as 1.0 × 10−5–4.5 × 10−5 M with a low limit of detection (LOD) (7.51 μM). Antimicrobial properties of cel-based compounds and Ag(I)-containing compounds were tested against five bacteria; Bacillus cereus, Streptococcus mutans, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and two fungi; Candida albicans and Candida tropicalis. The materials exhibited antimicrobial effects and their antifungal properties were more effective than their antibacterial properties.