Objectives: Ex vivo confocal laser scanning microscopy (EVCM) is a pioneering diagnostic method that enables fresh tissue samples to be analyzed directly during surgery. For the assessment of non-melanocytic skin cancer (NMSC), including cutaneous squamous cell carcinoma (cSCC), it provides a rapid addition to conventional histology. While previous studies have shown that EVCM reliably identifies the morphological features of cSCCs, quantitative criteria such as tumor thickness have not yet been systematically evaluated. This study investigated whether EVCM can be used to accurately and reproducibly measure the thickness of cSCCs, an important parameter for predicting metastatic risk.Methods: Eighty-two histologically verified cSCCs from different anatomical sites were assessed by the current gold standard of histopathology and EVCM. A statistical comparison of the confocal tumor thickness (CTT) and the histopathological tumor thickness (HTT) was then performed. In addition, it was analyzed how reliable EVCM was in the assignment of cSCCs to the correct tumor thickness category.Results: There was a very high agreement between both methods, evidenced by a Spearman correlation coefficient of 0.94 and a coefficient of determination of 0.859. Overall, 95.1% of the samples were correctly classified into the appropriate tumor thickness category using EVCM. Cohen’s Kappa of 0.90 indicated almost perfect agreement between EVCM and histology.Conclusions: These findings demonstrate that EVCM is a precise and reliable method for determining tumor thickness and the corresponding category in cSCCs. It enables immediate intraoperative assessment of the metastatic risk and preliminary classification of low-risk tumors. Additional studies with larger patient cohorts are required to further validate these results and support clinical implementation.
目的：离体共聚焦激光扫描显微镜（EVCM）是一种创新的诊断方法，可在手术过程中直接对新鲜组织样本进行分析。对于非黑色素细胞性皮肤癌（NMSC），包括皮肤鳞状细胞癌（cSCC）的评估，该方法为传统组织学提供了快速的补充。尽管先前研究表明EVCM能可靠识别cSCC的形态学特征，但肿瘤厚度等定量标准尚未得到系统评估。本研究旨在探讨EVCM能否准确且可重复地测量cSCC的厚度——这是预测转移风险的重要参数。
方法：通过现行金标准组织病理学与EVCM对来自不同解剖部位的82例经组织学验证的cSCC进行评估。随后对共聚焦肿瘤厚度（CTT）与组织病理学肿瘤厚度（HTT）进行统计学比较。此外，还分析了EVCM在将cSCC归入正确肿瘤厚度分类方面的可靠性。
结果：两种方法间具有高度一致性，斯皮尔曼相关系数为0.94，决定系数为0.859。总体而言，使用EVCM有95.1%的样本被正确分类至相应的肿瘤厚度类别。科恩卡帕值为0.90，表明EVCM与组织学评估几乎完全一致。
结论：这些发现证明EVCM是确定cSCC肿瘤厚度及相应分类的精确可靠方法。该方法能实现术中即时评估转移风险，并对低风险肿瘤进行初步分类。未来需要通过更大患者队列的研究进一步验证这些结果，以支持临床推广应用。
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