Objective To explore the clinical value of high-frequency ultrasound-guided labial gland biopsy in patients with suspected Sjögren’s syndrome (SS). Methods Totally 126 patients with clinically suspected SS and salivary gland involvement who underwent labial gland biopsy in Department of Ultrasound and Department of Rheumatology and Immunology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from Sep. 2022 to Dec. 2024 were enrolled. The patients who underwent traditional excision labial gland biopsy from Sep. 2022 to Feb. 2023 were selected as control group (n＝53), and the patients who received high-frequency ultrasound-guided labial gland biopsy from Mar. 2023 to Dec. 2024 were selected as ultrasound-guided group (n＝73). The success rate of sampling, the number of salivary lobes, wound healing time, and adverse reaction incidence were analyzed in the 2 groups. Results The success rate of sampling in the ultrasound-guided group (100.00%, 73/73) was significantly higher than that in the control group (92.45%, 49/53), the number of salivary lobes (6 ［6, 7］) was significantly greater than that in the control group (6 ［5, 6］), and the wound healing time was significantly shorter than that in the control group (4 ［4, 5］ d vs 5 ［5, 6］ d) (all P＜0.05). One (1.37%) patient of the ultrasound-guided group had postoperative adverse reactions, and 5 (9.43%) patients of the control group had adverse reactions, with no significant difference (P＞0.05). Conclusion High-frequency ultrasound can clearly locate labial gland, improve the success rate of sampling and shorten the wound healing time, and it is worthy of clinical promotion and application.