目的本研究加工了一种封接有多孔聚碳酸酯薄膜的过滤式微流控芯片用于循环肿瘤细胞分选。方法依据肿瘤细胞与血细胞在粒径和变形能力方面的差别,这种微过滤式芯片可以选择性截留肿瘤细胞。循环肿瘤细胞分析整个过程,包括细胞过滤、固定、标记和计数,均在芯片上完成。使用8μm孔径滤膜,可以选择性截留Hela细胞。以Hela细胞为模型,实验优化了肿瘤细胞的分选条件并考查了分选效果。结果在500µL/min流速下,Hela细胞的回收率可以达到85%,而血细胞残留可以控制在3000个以下。截留细胞使用微小染色体维持蛋白7抗体标记,可以有效辨别肿瘤细胞与血细胞。结论基于过滤式微流控芯片上的循环肿瘤细胞分选方法简单、快速、廉价,有望成为一种实用的循环肿瘤细胞检测技术。% Objective We fabricated a microfluidic device with embedded porous film for circulating tumor cell (CTC) enrichment from blood. Methods The microfilter can separate tumor cells from whole blood on the basis of differences in the size and deformability between tumor and hematologic cells. All the steps related to CTC analysis, including cell filtration, fixation, labeling and enumeration, were integrated on the same device. Our results demonstrated that Hela cells can specifically trapped on the porous film with 8μm pore-size. Using Hela cell as a model, operation conditions of the microfilter assay were optimized. Results At 500 µL/min flow rate, the capturing efficiency of Hela cell was 85%and the number of unspecific trapped hematologic cells can be limited to~3000 per test. Conclusions Using minichromosome maintenance protein 7 as the CTC characterization marker, the trapped Hela cells can be distinguished from the background blood cells. The microfilter-based CTC detection assay proposed here is simple, fast and low-cost, thus hold the potential to be developed into a practical CTC detection tool.
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