Almeida-Magana Ricardo, Au Matthew, Al-Hammouri Tarek, Mathew Manju, Dinneen Kate, Mendes Larissa S T, Dinneen Eoin, Vreuls Willem, Shaw Greg, Freeman Alex, Haider Aiman
Department of Targeted Intervention, University College, London, UK.
Department of Urology, University College London Hospitals NHS Foundation Trust, London, UK.
Histopathology. 2025 Feb;86(3):433-440. doi: 10.1111/his.15336. Epub 2024 Oct 15.
Fluorescence confocal microscopy (FCM) is a new imaging modality capable of generating digital microscopic resolution scans of fresh surgical specimens, and holds potential as an alternative to frozen section (FS) analysis for intra-operative assessment of surgical margins. Previously, we described the LaserSAFE technique as an application of FCM for margin assessment in robot-assisted radical prostatectomy (RARP) using the Histolog® scanner. This study describes the accuracy and inter-rater agreement of FCM imaging compared to corresponding paraffin-embedded analysis (PA) among four blinded pathologists for the presence of positive surgical margins (PSM).
RARP specimens from patients enrolled in the control arm of the NeuroSAFE PROOF study (NCT03317990) were analysed from April 2022 to February 2023. Prostate specimens were imaged using the Histolog® scanner before formalin fixation and PA. Four trained assessors, blinded to PA, reviewed and analysed FCM images of the posterolateral prostatic surface.
A total of 31 prostate specimens were included in the study. PA per lateral side of the prostate identified 11 instances of positive margins. Among the four histopathologists included in our study, FCM achieved a sensitivity of 73-91 and specificity of 94-100% for the presence of PSM. Fleiss' Kappa for inter-rater agreement on PSM was 0.78 (95% confidence interval = 0.64-0.92), indicating substantial agreement.
This blinded analysis of FCM versus PA among histopathologists with different experience levels demonstrated high accuracy and substantial inter-rater agreement for diagnosing PSM. This supports the role of the FCM as an alternative to FS.
荧光共聚焦显微镜（FCM）是一种新的成像方式，能够对新鲜手术标本进行数字显微分辨率扫描，并且作为术中评估手术切缘的冰冻切片（FS）分析的替代方法具有潜力。此前，我们将LaserSAFE技术描述为使用Histolog®扫描仪的FCM在机器人辅助根治性前列腺切除术（RARP）中评估切缘的一种应用。本研究描述了在四位不知情的病理学家之间，FCM成像与相应石蜡包埋分析（PA）相比，对于阳性手术切缘（PSM）存在情况的准确性和评估者间一致性。
2022年4月至2023年2月期间，对参与NeuroSAFE PROOF研究（NCT03317990）对照臂的患者的RARP标本进行分析。前列腺标本在福尔马林固定和PA之前使用Histolog®扫描仪进行成像。四位经过培训的评估者在不知情PA结果的情况下，对前列腺后外侧表面的FCM图像进行了审查和分析。
本研究共纳入了31个前列腺标本。前列腺每一侧的PA识别出11例阳性切缘。在我们研究的四位组织病理学家中，FCM对于PSM存在情况的敏感性为73 - 91%，特异性为94 - 100%。评估者间关于PSM的一致性的Fleiss' Kappa为0.78（95%置信区间 = 0.64 - 0.92），表明有实质性一致性。
在不同经验水平的组织病理学家中对FCM与PA进行的这项盲法分析表明，在诊断PSM方面具有高准确性和评估者间的实质性一致性。这支持了FCM作为FS替代方法的作用。
