Quantification of tumour vascularity in squamous cell carcinoma of the tongue using CARD amplification, a systematic sampling technique, and true colour image analysis.
Quantification of tumour vascularity in squamous cell carcinoma of the tongue using CARD amplification, a systematic sampling technique, and true colour image analysis.
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DOI：
10.1155/2001/780576
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
de Wilde PC
中科院分区：
文献类型：
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作者：
Hannen EJ;van der Laak JA;Kerstens HM;Cuijpers VM;Hanselaar AG;Manni JJ;de Wilde PC
这项头颈组织样本研究的目的是开发一种基于催化报告沉积 (CARD) 技术的免疫组织化学方案，以增强用于自动真彩色图像分析的染色结果，评估血管生成热点选择中系统组织采样的再现性，以及微血管密度 (MVD) 和其他血管特征的量化。后者的数据在六例转移性舌鳞状细胞癌与四例非转移性舌鳞状细胞癌之间进行了比较。与使用抗 CD34 抗体的标准免疫组织化学方案相比，CARD 扩增导致染色更强烈，血管数量更多。对组织切片中 40 至 60 个系统采样区域中的 10 个血管化程度最高的区域进行平均，得出大多数评估血管参数的总体可接受的观察者间再现性（r ≥ 0.76 且 p ≤ 0.01）。在非转移性舌癌中，直径 <5 μm 的血管百分比显着更高 (p = 0.02)。然而，对于许多肿瘤来说，组织取样的效果是显着的。我们的结论是，图像分析系统需要 CARD 放大来可靠地分割血管，并且肿瘤异质性是在单个组织切片中评估肿瘤血管分布的所有程序的限制因素。
The aims of this study of head and neck tissue samples were to develop an immunohistochemical protocol based on the catalysed reporter deposition (CARD) technique to enhance staining results for use in automated true colour image analysis, to assess the reproducibility of systematic tissue sampling in the angiogenic hot spot selection, and quantification of microvessel density (MVD) and other vessel characteristics. The latter data were compared between six metastasised tongue squamous cell carcinomas, vs. four non-metastasised. In comparison to the standard immunohistochemical protocol with anti-CD34 antibodies, CARD amplification resulted in both more intensely stained and larger numbers of vessels. Averaging the 10 most vascularised fields of the 40 to 60 systematically sampled fields in a tissue section resulted in an overall acceptable interobserver reproducibility for most assessed vessel parameters (r ≧ 0.76 and p ≦ 0.01). The percentage vessels with diameter <5 μm was significantly higher in the non-metastasised tongue carcinomas (p = 0.02). However, for a number of tumours the effect of tissue sampling was significant. We conclude that CARD amplification is needed for reliable segmentation of vessels by image analysis systems, and that tumour heterogeneity is a limiting factor for all procedures in which tumour vascularity is assessed in a single tissue section.