Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore138669.
Lab Chip. 2012 Nov 7;12(21):4388-96. doi: 10.1039/c2lc20750h.
Herein we present a lab-chip device for highly efficient and rapid detection of circulating tumor cells (CTCs) from whole blood samples. The device utilizes a microfabricated silicon microsieve with a densely packed pore array (10(5) pores per device) to rapidly separate tumor cells from whole blood, utilizing the size and deformability differences between the CTCs and normal blood cells. The whole process, including tumor cell capture, antibody staining, removal of unwanted contaminants and immunofluorescence imaging, was performed directly on the microsieve within an integrated microfluidic unit, interconnected to a peristaltic pump for fluid regulation and a fluorescence microscope for cell counting. The latter was equipped with a dedicated digital image processing program which was developed to automatically categorize the captured cells based on the immunofluorescence images. A high recovery rate of >80% was achieved with defined numbers of MCF-7 and HepG2 cancer cells spiked into human whole blood and filtered at a rapid flow rate of 1 mL min(-1). The device was further validated with blood drawn from various cancer patients (8 samples). The whole process, from sample input to result, was completed in 1.5 h. In addition, we have also successfully demonstrated on-microsieve fluorescence in situ hybridization for single cell molecular analysis. This simple method has great potential to supplant existing complex CTC detection schemes for cancer metastasis analysis.
在这里，我们提出了一种用于从全血样本中高效快速检测循环肿瘤细胞（CTC）的芯片装置。该装置利用具有密集孔阵列的微加工硅微筛（每个装置有 10^5 个孔），利用 CTC 和正常血细胞之间的大小和变形性差异，快速分离肿瘤细胞。整个过程，包括肿瘤细胞捕获、抗体染色、去除不需要的污染物和免疫荧光成像，都直接在集成微流控单元内的微筛上进行，与蠕动泵相连以进行流体调节，与荧光显微镜相连以进行细胞计数。后者配备了专门的数字图像处理程序，该程序用于根据免疫荧光图像自动对捕获的细胞进行分类。将 MCF-7 和 HepG2 癌细胞以预定数量混入人全血中，并以 1 mL min^(-1)的快速流速过滤，可获得>80%的高回收率。该装置还通过来自各种癌症患者的血液（8 个样本）进行了验证。从样本输入到结果，整个过程在 1.5 小时内完成。此外，我们还成功地在微筛上进行了荧光原位杂交，用于单细胞分子分析。这种简单的方法很有潜力取代现有的复杂 CTC 检测方案，用于癌症转移分析。
