Developing multiple concentration gradients for single cell–level drug screening NatRevDrugDi
was used for the fluids in the simulations. In addition, for fluid mixing calculation, water A and B are set as 0 and 1, respectively. A second-order limiting scheme was used to solve the species diffusion. The convergence limit for the mass fraction was set to 10Experimental setupSupplementary Information
. Three Longer® syringe pumps were utilized to regulate the operated flow rates. A 25 cm long Tygon soft tubing with an inner diameter of 0.42 mm was used for connection. Prior to use, the microchip was initially irradiated with UV light for 2 h and then sequentially rinsed with 70% ethanol for 2–3 min to eliminate air bubbles at a flow rate of 10 μL ∙ min, followed by rinsing in ultrapurified water and PBS working buffer.
To assess the efficacy of 5-fluorouracil and cisplatin in our cell culture models, cells were plated and grown to 50–70% confluence prior to treatment with 5-FU and DDP at increasing concentrations that are in accordance with concentrations formed by the microfluidic device. Subsequent to treatment, cells were tested for the efficacy of 5-FU and DDP, as described below. All cells were cultured for ≤ 4 months before being discarded, and fresh, frozen cells were used to continue studies.
For single-cell capture devices in the microfluidic chip, the devices were initially sterilized using sequential rinsing, i.e., by initially flushing 75% ethanol, followed by Millipore ultrapure water, PBS and fresh DMEM and then treated with a sterilized 50 μg ∙ mlfibronectin solution for 2 h at 37 °C to promote cell adherence, followed by cell seeding at a cell density of 1.0 × 10at a flow rate of 200 μL/min.
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