Karen J K, Gareau D S, Dusza S W, Tudisco M, Rajadhyaksha M, Nehal K S
Memorial Sloan-Kettering Cancer Center, New York, NY 10022, USA.
Br J Dermatol. 2009 Jun;160(6):1242-50. doi: 10.1111/j.1365-2133.2009.09141.x. Epub 2009 Mar 30.
High-resolution real-time imaging of human skin is possible with a confocal microscope either in vivo or in freshly excised tissue ex vivo. Nuclear and cellular morphology is observed in thin optical sections, similar to that in conventional histology. Contrast agents such as acridine orange in fluorescence and acetic acid in reflectance have been used in ex vivo imaging to enhance nuclear contrast.
To evaluate the sensitivity and specificity of ex vivo real-time imaging with fluorescence confocal mosaicing microscopy, using acridine orange, for the detection of residual basal cell carcinoma (BCC) in Mohs fresh tissue excisions.
Forty-eight discarded skin excisions were collected following completion of Mohs surgery, consisting of excisions with and without residual BCC of all major subtypes. The tissue was stained with acridine orange and imaged with a fluorescent confocal mosaicing microscope. Confocal mosaics were matched to the corresponding haematoxylin and eosin-stained Mohs frozen sections. Each mosaic was divided into subsections, resulting in 149 submosaics for study. Two Mohs surgeons, who were blinded to the cases, independently assessed confocal submosaics and recorded the presence or absence of BCC, location, and histological subtype(s). Assessment of confocal mosaics was by comparison with corresponding Mohs surgery maps.
The overall sensitivity and specificity of detecting residual BCC was 96.6% and 89.2%, respectively. The positive predictive value was 92.3% and the negative predictive value 94.7%. Very good correlation was observed between confocal mosaics and matched Mohs frozen sections for benign and malignant skin structures, overall tumour burden and location, and identification of all major histological subtypes of BCC.
Fluorescent confocal mosaicing microscopy using acridine orange enables detection of residual BCC of all subtypes in Mohs fresh tissue excisions with high accuracy. This observation is an important step towards the long-term clinical goal of using a noninvasive imaging modality for potential real-time surgical pathology-at-the-bedside for skin and other tissues.
使用共聚焦显微镜可在体内或体外新鲜切除的组织中对人体皮肤进行高分辨率实时成像。在薄光学切片中可观察到细胞核和细胞形态，这与传统组织学类似。荧光中的吖啶橙和反射率中的乙酸等造影剂已用于体外成像以增强核对比度。
评估使用吖啶橙的荧光共聚焦拼接显微镜进行体外实时成像检测莫氏新鲜组织切除标本中残留基底细胞癌（BCC）的敏感性和特异性。
在莫氏手术完成后收集48个废弃的皮肤切除标本，包括有和无所有主要亚型残留BCC的切除标本。组织用吖啶橙染色并用荧光共聚焦拼接显微镜成像。共聚焦拼接图像与相应的苏木精和伊红染色的莫氏冰冻切片匹配。每个拼接图像被分成子区域，共得到149个用于研究的子拼接图像。两名对病例不知情的莫氏外科医生独立评估共聚焦子拼接图像，并记录BCC的有无、位置和组织学亚型。通过与相应的莫氏手术图谱比较来评估共聚焦拼接图像。
检测残留BCC的总体敏感性和特异性分别为96.6%和89.2%。阳性预测值为92.3%，阴性预测值为94.7%。在良性和恶性皮肤结构、总体肿瘤负荷和位置以及BCC所有主要组织学亚型的识别方面，共聚焦拼接图像与匹配的莫氏冰冻切片之间观察到非常好的相关性。
使用吖啶橙的荧光共聚焦拼接显微镜能够高精度地检测莫氏新鲜组织切除标本中所有亚型的残留BCC。这一观察结果是朝着使用非侵入性成像模式实现皮肤和其他组织潜在的床边实时手术病理学这一长期临床目标迈出的重要一步。
