Background <p>The practice of endoscopic ultrasound-guided liver biopsy (EUS-LB) is becoming more common due to its proven safety and effectiveness. For accurate diagnosis, it is vital to secure ample tissue specimens. However, gauging the volume of tissue specimens accurately poses a challenge with existing methods. Additionally, determining the most suitable fine-needle biopsy (FNB) needle requires further study. Our aim was to contrast the tissue surface areas obtained using Franseen and Fork-tip needles and to identify factors affecting tissue volume.</p> Methods <p>This retrospective study analyzed liver tissue samples collected through EUS-LB using 19-gauge Franseen and Fork-tip needles from patients suffering from diffuse liver diseases, conducted in our hospital from April 2019 to April 2022. We primarily focused on measuring hepatic tissue surface area and portal tract count, alongside examining patient-related factors that could influence tissue surface area.</p> Results <p>The study involved 20 cases for each type of needle. The comparison revealed no significant disparities in the total liver tissue surface area (22.0 mm² vs. 22.6 mm², <i>P</i> = 0.45) or in the portal tract counts (30 vs. 20, <i>P</i> = 0.16). No adverse incidents were noted in either group. Both univariate and multivariate analyses highlighted that fibrosis and metabolic dysfunction associated steatotic liver disease (MASLD) presence were significant determinants of the total hepatic tissue area (<i>P</i> = 0.04, <i>P</i> &lt; 0.05; and <i>P</i> = 0.02, <i>P</i> = 0.03, respectively).</p> Conclusion <p>The capabilities of both needles in acquiring liver tissue were comparably effective. The volume of tissue was affected by the severity of fibrosis and the occurrence of MASLD.</p>