Iwamoto S, Burrows R C, Born D E, Piepkorn M, Bothwell M
Department of Physiology and Biophysics, University of Washington, Seattle 98195, USA.
Biomol Eng. 2000 Oct;17(1):17-22. doi: 10.1016/s1389-0344(00)00060-5.
A diagnostic problem can occur at the time of intraoperative consultation of neurosurgical tumors as to whether the tumor is of neuroectodermal origin or whether it represents an epithelial metastasis from another site. Intraoperative diagnoses based on hematoxylin and eosin stained frozen sections are often later confirmed by immunocytochemical analysis of formalin-fixed, paraffin-embedded tissue sections that are not available at the time of surgery. The objective of the current study was to demonstrate that the application of direct immunofluorescence to the intraoperative diagnosis of neurosurgical tumors would provide unequivocal, and nearly immediate results. This report describes a new application of an existing technique for an optimized, rapid procedure utilizing direct immunocytochemistry with fluorescence-labeled primary antibodies to analyze surgical biopsies intraoperatively. The examination of five neurosurgical biopsies established a neuroectodermal origin of three tumors via immunolabeling for glial fibrillary acidic protein (GFAP) and lack of labeling with keratin markers, whereas several metastatic lung carcinomas were identified by immunostaining for keratin, but not GFAP, markers. The results of the direct immunolabeling method were unequivocal and required only minutes. The same diagnoses were confirmed by standard immunocytochemical labeling of formalin-fixed, paraffin-embedded sections, though it required several days to obtain the results. Direct immunofluorescence using fluorescently conjugated primary antibodies is a practical and rapid method for deciding whether a neurosurgical tumor is a primary glial or an epithelial metastatic tumor in origin. It is the first reported application of the technique for this aspect of rapid neurosurgical diagnosis.
在神经外科肿瘤的术中会诊时，可能会出现诊断问题，即肿瘤是神经外胚层起源还是代表来自其他部位的上皮转移瘤。基于苏木精和伊红染色的冰冻切片的术中诊断，通常随后会通过对手术时无法获得的福尔马林固定、石蜡包埋组织切片进行免疫细胞化学分析来证实。本研究的目的是证明将直接免疫荧光应用于神经外科肿瘤的术中诊断将提供明确且几乎即时的结果。本报告描述了一种现有技术的新应用，即利用荧光标记的一抗进行直接免疫细胞化学，对手术活检组织进行优化的快速程序术中分析。对五例神经外科活检组织的检查通过对胶质纤维酸性蛋白（GFAP）进行免疫标记且角蛋白标记物未标记，确定了三例肿瘤的神经外胚层起源，而通过对角蛋白而非GFAP标记物进行免疫染色鉴定出了几例转移性肺癌。直接免疫标记法的结果明确，仅需几分钟。通过对福尔马林固定、石蜡包埋切片进行标准免疫细胞化学标记也证实了相同的诊断，不过获得结果需要几天时间。使用荧光偶联一抗的直接免疫荧光是一种实用且快速的方法，用于确定神经外科肿瘤是原发性神经胶质肿瘤还是上皮转移性肿瘤。这是该技术首次报道用于快速神经外科诊断的这一方面。
