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Model pdc 001

Manufactured by Harrick

The Harrick PDC-001 is a high-performance plasma cleaner. It is designed to clean and activate surfaces using a low-pressure oxygen plasma. The device operates at a frequency of 13.56 MHz and can accommodate samples up to 4 inches in diameter.

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4 protocols using model pdc 001

1

Coverslip Preparation and Flow Chamber Construction

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Coverslips (24 mm × 60 mm; VWR, #16004-312) were cleaned by sonicating in distilled water for 10 min, followed by sonicating in acetone for 10 min. After air-drying in a hood, the coverslips were plasma cleaned in argon for 5 min on the high setting (Harrick Plasma, Model PDC-001). The plasma-cleaned slides were mounted in home-machined aluminum microscope holders and held in place by high-vacuum grease (Dow Corning, VWR #59344-055). Flow chambers were constructed from strips of double-sided tape and a top coverslip (10.5 mm × 22 mm; Electron Microscopy Sciences, #72191-22).
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2

Electrospun CSS-PCL Composite Fibers

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CSS was synthesized, according to a procedure detailed in our previous studies [7 (link),38 (link)]. An acidic solution of CSS at a concentration of 69% w/w was prepared in a mixed solvent that comprises 1 mL of tetrahydrofuran (Sigma Aldrich) and 10μL of 37% aqueous HCl. The solution was incubated overnight in a 40°C water bath, permitting CSS hydrolysis and polymerization. The hydrolyzed and polymerized CSS solution was then electrospun into lipid fibrous membranes using a custom made device with a flow rate of 0.5 μL/min, a voltage of 12 kV, and a spinneret-to-ground distance of 12 cm. The fibers were collected on silicon chips.
A solution of PCL (MW: 80 kDa, Sigma Aldrich) at a concentration of 10% w/v was prepared in 1,1,1,3,3,3-Hexafluoro-2-propanol (MW: 168.04, Sigma Aldrich). The solution was incubated at room temperature for a minimum of 6 h, briefly vortexed, and then electrospun into fiber with a flow rate of 20 μL/min, a voltage of 12 kV, and a spinneret-to-ground distance of 12 cm. The electrospun PCL fibers were collected on silicon chips that were placed on top of the aluminum foil collector plate. Approximately half of the collected PCL fibers immediately underwent an air-plasma treatment (Harrick Plasma, Model PDC-001) for 10 min under vacuum, generating plasma-treated PCL fibers.
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3

Fabrication of PDMS Microfluidic Devices

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Polyurethane-based plastic (Smooth-Cast ONYX SLOW, Smooth-On) was used to make replicas of the microstructures on the master silicon wafer, as previously described45 (link). PDMS microfluidic devices were then fabricated from these molds using soft-lithography of RTV 615 PDMS (Momentive Performance Materials).
After baking, the cured microfluidic device was removed from its mold, and holes for the fluidic introduction ports including cross flow inlet (CFI) and sample inlet (SI) as well as the oscillation inlets (OSC1 and OSC2), were punched into it using a 0.5 mm outer diameter hole punch (Technical Innovations, Angleton, TX, USA). The outlets were punched using a 3 or 4 mm diameter puncher. The microfluidic device was then bonded by oxygen plasma (Model PDC-001, Harrick Plasma) to a blank layer of PDMS previously spin-coated onto a blank silicon wafer at 1500 rpm for 1 minute and then subsequently bonded to a standard microscope slide (50 × 75 mm, Fisher Scientific).
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4

Microfluidic Device Fabrication and Cell Adhesion

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The microfluidic device was comprised of a molded poly(dimethylsiloxane) (PDMS, General Electric RTV 650 Part A/B) slab bonded to a glass substrate. High-resolution printing (5080 dpi) was used to print the mask with the design pattern on a transparency film. The mask was used to fabricate 50 µm high SU-8 2050 photoresist (Microchem) features on a silicon wafer via photolithography. PDMS molds with embossed channels were fabricated using soft lithography by curing the pre-polymer on the silicon master for 2 hours at 70°C. The PDMS replica was then peeled off the silicon master. Inlets and outlets for the fluids and cells were created in PDMS using a steel punch. The surface of the PDMS replica and a clean glass coverslip (Fisher Scientific) were treated with air plasma for 90 seconds (Model PDC-001, Harrick Scientific) and irreversibly bonded to complete the device assembly (Figure S7). The device inlets were then connected to 1 mL syringes (BD Biosciences) with 23 G ¾ size needles (BD Biosciences) via PTFE tubing (Cole-Parmer). All syringes were calibrated and pushed by a constant pressure syringe pump (Harvard Apparatus). Prior to each experiment, the device was also loaded with fibronectin (25 µg/mL, Invitrogen) and kept at room temperature for 30 minutes to promote optimal cell adhesion.
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