Design and fabrication of a microchip. (a) Principle of selective cell capture using antibodies. Shallow channel depth is designed to be slightly smaller than the cells. Cells are captured in this part. (b) The fabricated microchip (70 mm× 30 mm). (c) The microchannel around the shallow channel (4× magnification).
Scheme of in situ RCA on captured cells. (a) Detection of mitochondrial DNA in HL-60 cells. (b) Detection of mRNA (HER2) in SK-BR-3 cells.
HL-60 cell capture and RCA. (a) Captured HL-60 cells in a shallow channel. (b) Capture rate versus flow velocity in a shallow channel at channel depths of 8 μm and 10 μm.
RCA detection of mitochondrial DNA in captured HL-60 cells. (a) Bright field image. (b) Fluorescence image. (c)Dots per cell versus padlock probe concentration. Values are represented as mean ± SD; n = 50.
Fluorescence images of a microchannel with and without antibody coating. (a) An antibody-coated microchannel. (b) A non-coated microchannel.
Cell capture in an antibody-coated shallow microchannel. (a) Cell selectivity under various conditions. Condition A included SK-BR-3 cells and a non-coated microchannel. Condition B included RPMI-1788 cells and an antibody-coated microchannel. Condition C included SK-BR-3 cells and an antibody-coated microchannel. In all conditions, channel depth was 16 μm. The diameter of SK-BR-3 cells and RPMI-1788 cells is 12–24 μm and 16 μm, respectively. (b) Capture rate versus flow velocity in each condition. (c) Captured SK-BR-3 cells in condition C.
RCA detection of HER2 in captured SK-BR-3 cells. (a) Bright field. (b) A fluorescence image.
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