BACKGROUND: Early detection and treatment are critical for improving the survival and prognosis of patients with cervical cancer. However, there is a notable scarcity of targeted imaging probes specifically designed to detect high-grade squamous intraepithelial lesions (HSIL) and cervical cancer. Our study aimed to address this gap by identifying and validating a targeted imaging probe for these conditions. RESULTS: Using bioinformatics data, we identified galectin-7 (GAL7) as highly expressed in patients with cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC). Immunohistochemical staining of biopsy samples from 30 HSIL and cervical cancer patients verified the high and specific expression of GAL7. Further validation was performed using mouse and human CESC cell lines and tumor xenografts, confirming the consistent expression of GAL7. Based on this finding, we synthesized a GAL7-specific antibody conjugated with FITC, creating the GAL7-FITC fluorescence imaging probe. Fluorescence molecular imaging revealed that GAL7-FITC exhibited specific binding to various CESC cell lines and xenograft mouse models. Additionally, the diagnostic capability of GAL7-FITC was demonstrated in fresh HSIL specimens from cervical cone excisions, validated through histopathology and immunohistochemical analysis. CONCLUSIONS: Our study identified GAL7 as a specific target for CESC and successfully developed the GAL7-FITC fluorescence imaging probe. GAL7-FITC has shown promising potential for clinical application in the early detection of HSIL and CESC, providing rapid fluorescence imaging diagnosis without observable toxicity. This advancement may significantly enhance the accuracy and speed of cervical cancer diagnostics, ultimately improving patient outcomes.