- 新一代离体荧光共聚焦显微镜（EVFCM）以「数字染色」模式生成类 HE 图像；作者在 39 例患者的 42 个非黑色素瘤皮肤癌（NMSC）Mohs 手术标本中前瞻性评估其切缘判读，共 261 个切缘。
- 皮肤科医生与病理医生（均受过 EVFCM 判读训练）独立盲读：切缘判读 κ=0.863（观察一致率 98.6%），组织学类型判读 κ=0.882（97.6%）。
- 与常规组织病理对照，总体诊断准确率 96.4%，联合评估 AUC 0.875；仅 2 例不一致（1 例切缘假阴性、1 例源于样本处理差异）。流程时间优势仍需定量前瞻验证。
收录范围：PubMed 英文摘要及中文翻译（abstract-only）。Unpaywall 标注原文为 hybrid 开放获取（CC BY），但 ScienceDirect 与 modernpathology.org 对本站出口返回 Cloudflare 验证/登录页（curl 与 Chrome headless 均未通过），未采集全文、图表；全文待后续补采。
摘要
新一代离体荧光共聚焦显微镜（EVFCM）可生成与常规组织病理直接可比的类苏木精-伊红图像。其在非黑色素瘤皮肤癌（NMSC）显微描记控制手术中、采用一种新采集模式时的观察者间表现尚待明确。在这项前瞻性干预研究中，对 2025 年 1 月至 10 月间接受 Mohs 手术的 39 例患者的 42 个肿瘤进行了 EVFCM 检查。标本经正交切取、染色，并以数字染色模式采集图像。周边及基底切缘由一名皮肤科医生和一名病理医生独立、盲法判读，二人均接受过 EVFCM 判读培训。判读一致为阴性的切缘视为无肿瘤，而阳性、不一致或可疑的结果则需再次切除并重新评估。采用 Cohen κ 统计量评估与组织病理学的诊断一致性。共评估 261 个切缘。皮肤科医生与病理医生之间的判读者间一致性在切缘评估（κ=0.863；SE 0.067；p<0.001；观察一致率 98.6%）和组织学类型评估（κ=0.882；SE 0.031；p<0.001；观察一致率 97.6%）上均几乎完全一致。与组织病理学相比，EVFCM 表现优良，总体诊断准确率为 96.4%。联合评估的 AUC 为 0.875（SE 0.074；95% CI 0.730–1.000）。EVFCM 与组织病理学之间仅观察到两处不一致：一处为切缘评估假阴性，另一处不一致归因于样本处理差异。这些发现支持 EVFCM 作为 NMSC 术中切缘评估的可靠工具。皮肤科医生与病理医生判读者之间相当的表现，支持其有望整合进 Mohs 手术工作流程；然而，标准化样本处理方法可能带来的流程优势，尚需采用定量时间指标的前瞻性验证。
- New-generation ex vivo fluorescence confocal microscopy (EVFCM) produces H&E-like images in digital-stain mode; the authors prospectively evaluated margin reading in 42 nonmelanoma skin cancers (NMSC) from 39 Mohs surgery patients — 261 margins.
- A dermatologist and a pathologist (both EVFCM-trained) read blindly and independently: κ=0.863 for margins (98.6% observed agreement) and κ=0.882 for histotype (97.6%).
- Against conventional histopathology, overall accuracy was 96.4% and AUC for the combined assessment 0.875; only two discrepancies (one false-negative margin, one due to sample-processing differences). Workflow time advantages still need quantitative prospective validation.
Scope: PubMed abstract only. Unpaywall lists the article as hybrid open access (CC BY), but ScienceDirect and modernpathology.org returned a Cloudflare challenge/login page to this site (curl and headless Chrome), so full text, figures and tables were not collected.
Abstract
New-generation ex vivo fluorescence confocal microscopy (EVFCM) generates hematoxylin-eosin-like images directly comparable to conventional histopathology. Its interobserver performance with a novel acquisition mode in non-melanoma skin cancer (NMSC) micrographically controlled surgery remains to be defined. In this prospective interventional study, 42 tumors from 39 patients undergoing Mohs surgery between January and October 2025 were examined with EVFCM. Specimens were orthogonally sectioned, stained, and acquired in digital-stain mode. Peripheral and deep margins were independently and blindly reviewed by a dermatologist and a pathologist, both trained in EVFCM interpretation. Concordant negative margins were considered tumor-free, whereas positive, discordant, or equivocal findings prompted re-excision and re-evaluation. Diagnostic agreement with histopathology was assessed using Cohen's κ statistic. A total of 261 margins were evaluated. Inter-reader agreement between the dermatologist and pathologist was almost perfect for both margin assessment (κ=0.863; SE 0.067; p<0.001; observed agreement 98.6%) and histotype assessment (κ=0.882; SE 0.031; p<0.001; observed agreement 97.6%). Compared with histopathology, EVFCM showed high performance, with an overall diagnostic accuracy of 96.4%. The AUC for the combined assessment was 0.875 (SE 0.074; 95% CI 0.730-1.000). Only two discrepancies were observed between EVFCM and histopathology: one false-negative margin assessment and one discordance attributable to differences in sample processing. These findings support EVFCM as a reliable tool for intraoperative margin assessment in NMSC. Comparable performance between dermatologist and pathologist readers supports its potential integration into Mohs surgery workflows; however, potential workflow advantages of the standardized sample-processing approach require prospective validation using quantitative time metrics.