Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06511, United States.
Nano Lett. 2012 Jun 13;12(6):2697-704. doi: 10.1021/nl2041707. Epub 2012 May 31.
We report on the development of a nanowire substrate-enabled laser scanning imaging cytometry for rare cell analysis in order to achieve quantitative, automated, and functional evaluation of circulating tumor cells. Immuno-functionalized nanowire arrays have been demonstrated as a superior material to capture rare cells from heterogeneous cell populations. The laser scanning cytometry method enables large-area, automated quantitation of captured cells and rapid evaluation of functional cellular parameters (e.g., size, shape, and signaling protein) at the single-cell level. This integrated platform was first tested for capture and quantitation of human lung carcinoma cells from a mixture of tumor cells and leukocytes. We further applied it to the analysis of rare tumor cells spiked in fresh human whole blood (several cells per mL) that emulate metastatic cancer patient blood and demonstrated the potential of this technology for analyzing circulating tumor cells in the clinical settings. Using a high-content image analysis algorithm, cellular morphometric parameters and fluorescence intensities can be rapidly quantitated in an automated, unbiased, and standardized manner. Together, this approach enables informative characterization of captured cells in situ and potentially allows for subclassification of circulating tumor cells, a key step toward the identification of true metastasis-initiating cells. Thus, this nanoenabled platform holds great potential for studying the biology of rare tumor cells and for differential diagnosis of cancer progression and metastasis.
我们报告了一种基于纳米线基底的激光扫描成像细胞仪的开发，用于稀有细胞分析，以实现对循环肿瘤细胞的定量、自动和功能评估。免疫功能化的纳米线阵列已被证明是一种从异质细胞群体中捕获稀有细胞的优异材料。激光扫描细胞仪方法能够在单细胞水平上实现大面积、自动定量捕获的细胞，并快速评估功能细胞参数（例如大小、形状和信号蛋白）。该集成平台首先用于从肿瘤细胞和白细胞混合物中捕获和定量人肺癌细胞进行了测试。我们进一步将其应用于在新鲜人全血中（每毫升几个细胞）中加入稀有肿瘤细胞的分析，这些全血模拟了转移性癌症患者的血液，并证明了该技术在临床环境中分析循环肿瘤细胞的潜力。使用高内涵图像分析算法，可以快速自动、无偏和标准化地定量细胞形态计量参数和荧光强度。总之，这种方法能够原位对捕获的细胞进行有意义的特征描述，并有可能对循环肿瘤细胞进行亚分类，这是鉴定真正的起始转移细胞的关键步骤。因此，这个纳米技术平台在研究稀有肿瘤细胞的生物学和癌症进展及转移的鉴别诊断方面具有很大的潜力。
