Overview of the CTC-Race assay and its device.
(a) Schematic of
CTC-Race assay. The microfluidic device comprises a cell-loading channel
(1), multiple single-cell racetrack segments (3), multiple cell-collection
channels (2), and a chemokine channel (4). The height of single-cell
racetracks is designed to be smaller than the diameter of cancer cells
so that cancer cells are trapped at the entrance of single-cell racetracks
at the start of the assay. Migratory cells (red) squeeze into the
tracks and migrate toward the region with a high concentration of
chemoattractants, while the nonmigratory cells (green) are trapped
in the cell-loading channel during the assay. Cell-collection channels
are inserted into the single-cell racetracks perpendicularly to enable
cell retrieval. (b) Photos of a PDMS-based CTC-Race device (right,
blue and orange colors indicate the channel geometry). (c) Top view
schematic of the CTC-Race device. Cells are injected via the cell-loading
inlets, and chemokines are continuously perfused through the chemokine
channel. The device has a symmetric design with each side having a
cell-loading channel (width of 200 μm, height of 50 μm),
three cell-collection channels (width of 100 μm, height of 50
μm), and four segments of single-cell racetracks (each segment’s
length, 150 μm, width of 30 μm, height of 5 μm).
There are 2500 single-cell racetracks on each side and 5000 racetracks
in total in one device. Both sides share a chemokine channel (width
of 200 μm, height of 50 μm). (d) Phase-contrast images
of the migration pattern of MDA-MB-231 breast cancer cells in the
CTC-Race device. Chemokine channel is loaded with serum (fetal bovine
serum, FBS) as a chemoattractant. (e) Top, side view of rhodamine
B (red) concentration at t = 0 h. Bottom left, a
fluorescent image of the device at t = 0 h of rhodamine
B perfusion with a flow rate of 0.1 μL/min. Bottom right, normalized
fluorescence intensity across the device at the selected positions
in the device (yellow dots on the fluorescent image) at t = 0 h. (f) Top, side view of rhodamine B (red) at t = 24 h. Bottom left, a fluorescent image of the device at t = 24 h. Bottom right, normalized fluorescence intensity
across the device at t = 24 h.