Listeria monocytogenes can adapt, persist, and form biofilms on food premises surfaces, representing a challenge for food safety, since they led to disease transmission, food contamination and spoilage during production. Physical interventions (scrubbing and wiping) can help controlling formation, nevertheless when biofilms are formed, they are usually very resistant to current control strategies used in the food industry. Biofilm attachment and formation is influenced by environment characteristics, substrate properties and microbial motility. The purpose of this study was to evaluate the ability of L. monocytogenes to attach and form biofilms on different surfaces (wood, nylon, and polycarbonate) representative of the materials used during produce harvesting and storage. Multi-strain L. monocytogenes biofilms were grown in a CDC Biofilm reactor at 20 ± 2 °C up to 96-h and characterized for: a) attachment strength by enumerating cells after rinsing; b) hydrophobicity and interfacial tension by contact angle measurements; c) biofilm architecture by Laser Scanning Confocal Microscopy. All experiments were done in triplicate. Material, incubation, and solvent significantly affected the hydrophobicity and wetting properties of L. monocytogenes biofilms (P < 0.05). The type of material and incubation time significantly influenced hydrophobicity and wetting properties of L. monocytogenes biofilms (P < 0.05). Highest contact angle and lowest interfacial tension were observed on polycarbonate coupons. The data presented contributes to understanding Listeria biofilms grow on different surfaces commonly used in produce harvesting and storage. The data obtained in this study can be used when evaluating intervention strategies to control this pathogen in food premises.