Isolation and label-free detection of circulating tumour cells by fluidic diffraction chips with a reflective laser beam system

A photonic crystal (PC) based line array of poly(methacrylic acid) (PMAA) brushes was grafted from a photoresist template using a trench array. The array was functionalised with anti-epithelial cell adhesion molecule antibodies (EpY). A laser beam was employed to analyse the two-dimensional (2D) and three-dimensional (3D) reflective signals of PCs at an incidence angle of 45 degrees. The EpY-tailed PMAA PC possessed an optical feature with a characteristic diffraction effect along two laser input configurations including the SII configuration, in which the projection of the laser beam on the plane of the SPM chip was parallel to the strips, and the ST configuration, in which they were perpendicular. A fluidic diffraction chip based on the EpY-tailed PMAA PC with 1-mu m resolution was fabricated to examine the ability to detect circulating tumour cells (CTCs) along the ST configuration. The CTCs attached on the EpY-tailed PMAA PC, resulting in the change in the diffraction intensity. Dependence of change degree of the diffraction intensity exhibited a linear range of concentration of CTC from 0 to 64 cells and a limit of detection of 5 cells in 3 mL. CTC detection using both fluidic diffraction chips and a commercial IsoFlux system was carried out in clinical trials, including three healthy donors and 12 patients at various stages of colorectal cancer for comparison. Our platform provides a simple label-free method with high accuracy for rapid CTC counting, which has great potential in clinical treatment applications.