Microfluidic device for CTC detection, capture, and recovery. (a) Design of the chip. (b) Pre-filter with a gap space of 50 μm to minimize blood clogging. (c) Enlarged view of the cells capture region. The size distribution of the micro pillars varies from 12 μm down to 4 μm. (d) Details of the connection setup with a syringe pump to drive the cells through the microfluidic chip.
(a) Cell capture chamber depicting trapped Hela. (b) The DE ranges from a high of 95% to a low of 74% for the 4 flow rates (0.5, 1.0, 1.5, 2.0 ml/h) being studied. (c) A comparison of the 5-day cell proliferation process between captured cells (at the max flow rate of 2 ml/h) and normal cultured cells shows no perceptible difference between them. Thus, it appears that the viability of the cancer cells is unaffected by the 4 flow rates used for their capture. Further, our micropillars are designed with an unusual shape to minimize damage to a cell caught in-between 2 micropillars. Scale bar is 100 μm.
A comparison of the severity of blood clots in the capture region of the chip when working with (a) unlysed blood versus (b) lysed blood.
(a) Bright field and fluorescence and merged images in the microfluidic chip. Hela and WBCs are marked with red and green fluorescence, respectively. Scale bar is 10 μm. (b) The CP of the chip seems to be unaffected by the varying flow rates.
Positive CTC enumeration was detected in all blood samples taken from patients with liver cancer. (a) CTC count per 3-ml blood sample and it ranged from a low of 1–2 (1 patient) to a high of >20 (2 patients). (b) Captured cells from blood of liver cancer patients. Scale bar is 10 μm. (c) Immunofluorescence staining is performed on recovered CTCs. To distinguish the inadvertently captured WBCs from the captured CTCs, the former are stained with blue and red fluorescence, while the latter are stained with blue and green fluorescence. The merged slide clearly depicts the distinction between the 2 cell types. Scale bar is 10 μm.
Antibody-independent technology for CTC segregation, capture, and recovery.
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