A fluoropolymer membrane filter with through-holes was fabricated by photolithographic patterning and the dry etching method. 380,000 highly packed through-holes, each with a diameter of 7 µm were able to cover a whole area with a diameter of 13 mm. Ethylene tetrafluoroethylene (ETFE) was used as the membrane, which was suitable for the fluorescence detection of rare cells such as circulating tumor cells (CTCs) in human blood. The device fabrication for the size based capture of rare cells in blood such as CTCs is realized in this study.

Although membrane filters are indispensable in biochemical analysis fields, most methods for through-hole fabrication are complex and inefficient. We developed a simple method of fabricating poly(ethylene terephthalate) (PET) membrane filters with a precise arrangement of through-holes for the isolation of circulating tumor cells (CTCs) based on their size. By photolithography and dry etching, highly packed 380,000 through-holes with a diameter of 7 µm were able to cover a whole area with a diameter of 13 mm. Device fabrication for the size-based capture of rare cells in blood such as CTCs is realized in this study.

Considering the challenges in isolating circulating tumor cells (CTCs) pertaining to cellular stress and purity, we report the application of a blood microfiltration device as an optimal approach for noninvasive liquid biopsy to target CTCs. We experimentally analyzed the filtration behavior of the microfilter using pressure sensing to separate tumor cells from leukocytes in whole blood. This approach achieved an average recovery of >96% of the spiked tumor cells and depletion of >99% of total leukocytes. Furthermore, we carried out genomic profiling of the CTCs using the blood microfiltration device. The method was also applied in a clinical setting; DNA amplification was performed using a small number of microfiltered CTCs and epidermal growth factor receptor mutations were successfully detected to characterize the efficacy of molecularly targeted drugs against lung cancer. Overall, the proposed method can provide a tool for evaluating efficient filtration pressure to concentrate CTCs from whole blood.