BackgroundContrast-enhanced endoscopic ultrasound-guided fine needle aspiration (CE-EUS-FNA) could help clinicians to precisely locate and puncture lesions, but its effect on the diagnostic yield improvement is controversial. We designed this study to observe the additional benefit of using contrast in EUS-guided tissue sampling while performing fine needle biopsy (FNB) instead of FNA, as FNB results in a higher diagnostic accuracy.MethodPatients who underwent EUS-FNB performed by a single medical team from January 2019 to March 2021 were included in this study. We analyzed the cytopathological diagnostic accuracy rate and number of needle passes between groups who underwent FNB with and without contrast.ResultWe divided 133 patients who were diagnosed with a malignancy into two groups according to whether they underwent CE-EUS-FNB (n = 48) or conventional EUS-FNB (n = 85). The CE-EUS-FNB group had an equal diagnostic accuracy rate with fewer needle passes compared with the conventional EUS-FNB group. There was no significant trend change in the success cytopathological diagnostic rate for experienced endoscopists for EUS-FNA.ConclusionCE-EUS-FNB had fewer needle passes but no additional benefit for diagnostic yield improvement. There was no difficult threshold for CE-EUS-FNB for endoscopists who were well trained in conventional FNA.