RESUMEN
The fabrication of high-performance microscale devices in substrates with optimal material properties while keeping costs low and maintaining the flexibility to rapidly prototype new designs remains an ongoing challenge in the microfluidics field. To this end, we have fabricated a micro free-flow electrophoresis (µFFE) device in cyclic olefin copolymer (COC) via hot embossing using a PolyJet 3D-printed master mold. A room-temperature cyclohexane vapor bath was used to clarify the device and facilitate solvent-assisted thermal bonding to fully enclose the channels. Device profiling showed 55 µm deep channels with no detectable feature degradation due to solvent exposure. Baseline separation of fluorescein, rhodamine 110, and rhodamine 123, was achieved at 150 V. Limits of detection for these fluorophores were 2 nM, 1 nM, and 10 nM, respectively, and were comparable to previously reported values for glass and 3D-printed devices. Using PolyJet 3D printing in conjunction with hot embossing, the full design cycle, from initial design to production of fully functional COC µFFE devices, could be completed in as little as 6 days without the need for specialized clean room facilities. Replicate COC µFFE devices could be produced from an existing embossing mold in as little as two hours.
RESUMEN
During metastasis, cancer cells are exposed to potentially destructive hemodynamic forces including fluid shear stress (FSS) while en route to distant sites. However, prior work indicates that cancer cells are more resistant to brief pulses of high-level FSS in vitro relative to non-transformed epithelial cells. Herein, we identify a mechano-adaptive mechanism of FSS resistance in cancer cells. Our findings demonstrate that cancer cells activate RhoA in response to FSS, which protects them from FSS-induced plasma membrane damage. We show that cancer cells freshly isolated from mouse and human tumors are resistant to FSS, that formin and myosin II activity protects circulating tumor cells (CTCs) from destruction, and that short-term inhibition of myosin II delays metastasis in mouse models. Collectively, our data indicate that viable CTCs actively resist destruction by hemodynamic forces and are likely to be more mechanically robust than is commonly thought.