Aim: To analyze the impact of surface roughness on the visualization of latent fingerprint impressions. Methods: This study included eight adult participants who left their fingerprints on a polystyrene sheet. The sheet was treated with sandpaper of different grit sizes (60, 240, and 600 grit), while one remained untreated. After surface treatment, roughness measurements were performed on all surfaces. After the fingerprints were deposited, different powders were used for visualization: Magnetic Jet Black and Jet-Black Special powders. The prints were graded on a scale of 0–4. Descriptive statistical analysis, a linear mixed-effects model, and a mixed logistic regression model were used to analyze the effects of different factors on the quality of the visualized fingerprints. Results: Magnetic powder provided significantly better-quality fingerprints, with 96.9% being suitable for identification compared to 71.9% for the special powder. The magnetic powder exhibited better results on very smooth surfaces. The linear mixed-effects model revealed that increased surface roughness significantly decreased the quality of the fingerprints (P = 0.001), whereas the magnetic powder increased the fingerprint quality by 0.687 units (P < 0.001). The interaction between powder type and substrate showed that the magnetic powder yielded better results on rougher surfaces (P = 0.030). The mixed logistic regression model confirmed that magnetic powder increased the odds of a fingerprint being suitable for identification by 20.50 times (P = 0.011). Conclusion: Surface roughness significantly affects the visualization of latent fingerprint impressions and Magnetic Jet Black powder is recommended for use on rough surfaces. These findings are valuable for forensic experts, as they suggest that fingerprint quality can vary depending on the surface and powder used, which can affect their usability for identification purposes.