For CTC capture, PBS (200 μl) was first introduced into a Click Chip via a digital fluidic handler at a flow rate of 1 ml hour−1 to confirm that an appropriate seal was made between the patterned PDMS chaotic mixer and the Tz-grafted SiNWS. The TCO-grafted artificial CTC samples and peripheral blood mononuclear cell (PBMC) samples (200 μl) collected from patients with NSCLC were infused into Click Chip microfluidic devices at the optimum flow rate of 1 ml hour−1. For CTC enumeration, the CTCs captured in Click Chip were fixed with 2.0% formaldehyde (Electron Microscopy Sciences) in PBS (200 μl). CTCs captured on the chips were enumerated. To release CTCs, 200 μl of DTT (50 mM) was injected into the Click Chip system at a flow rate of 1 ml hour−1, and then 100 μl of PBS was injected at a flow rate of 1 ml hour−1 to collect the released CTCs. The released CTCs were collected into either a 96-well plate for fluorescence imaging or 1.5-ml ribonuclease (RNase)–free Eppendorf tubes for subsequent molecular analysis using RT-ddPCR.

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