We employed a custom microfluidic chip design with the architectures and operating mechanism described in Figure 2 . The fabrication for this four-nozzle droplet generation platform was based on a modified version of an established soft lithography procedure.24 (link) Microchannel architectures were designed using AutoCAD (Autodesk, San Rafael, CA, USA) and sent to CAD/ART services (Bandon, OR, USA) who provided high-resolution photomasks. SU-8 master molds were produced with the recommended protocol from Micro-Chem. After wafer fabrication, PDMS was mixed in a 1:10 ratio of curing agent to elastomer and subsequently degassed and poured onto the SU-8-on-Si wafer master. PDMS was cured in an oven for 4 hours at 95 °C. Once cured, PDMS layers were peeled off and punched with a 1 mm diameter biopsy punch (Kay Industries Co., Tokyo, Japan) to create inlets and outlets. To bond PDMS to glass, both surfaces were treated with oxygen plasma in a PDC-32G Harrick Plasma Cleaner (Harrick Plasma, Ithaca, NY) for 30 s and bound together. Finally, the droplet generation devices were post-baked in a convective oven at 95 °C for at least 24 hours.
Plasma cleaner pdc 32g
Manufactured by Harrick
Sourced in United States
The Plasma Cleaner PDC-32G is a laboratory equipment designed for surface cleaning and activation. It utilizes a plasma discharge to remove organic contaminants and enhance the surface properties of various materials.
Automatically generated - may contain errors
Lab products found in correlation
Glass slides, by Electron Microscopy Sciences (2 mentions)
Type 1 collagen, by BD (2 mentions)
Digital sight qi1mc, by Nikon (2 mentions)
Eclipse ti microscope, by Nikon (2 mentions)
2100 cryo tem, by JEOL (2 mentions)
Vitrobot mark 4, by Thermo Fisher Scientific (2 mentions)
Autocad, by Autodesk (1 mentions)
Sylgard 184, by Dow (1 mentions)
Cell tak adhesive, by Corning (1 mentions)
Sylgard 184 silicone elastomer, by Dow (1 mentions)
Microscope slide, by Thermo Fisher Scientific (1 mentions)
S 4800, by Hitachi (1 mentions)
Tecnai g2 f20 s twin, by Thermo Fisher Scientific (1 mentions)
Titan krios, by Ametek (1 mentions)
Au r1 2 1, by Quantifoil (1 mentions)
Droplet generation oil, by Bio-Rad (1 mentions)
Plain glass microscope slides, by Thermo Fisher Scientific (1 mentions)
35 protocols using plasma cleaner pdc 32g
1
Microfluidic Droplet Generation Platform
2
Hydrophobic and Hydrophilic Glass Treatments
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Hydrophobic treatment: We cleaned the glass slide use acetone, ethanol and deionized water in turn in an ultrasonic cleaning machine, then let the glass substrate dry naturally. The contact angle of the cleaned glass slide is 43°. Then we submerged the cleaned glass in the reactive solution for 60 s and withdrawned gradually. The substrate was allowed to dry about 5 min at room temperature (25 °C). Ethanol and water were used to rinse the glass surface. The contact angle of the hydrophobic substrate is 100o.
Hydrophilic treatment: We use oxygen plasma treatment (PDC-32G Harrick Plasma Cleaner) for the hydrophilic treatment of the glass substrate. Glass substrates were treated with low power oxygen plasma for 3 min, the contact angle is 10°. Glass substrates were treated with low power oxygen plasma for 15 min, and the contact angle is almost 0o.
Hydrophilic treatment: We use oxygen plasma treatment (PDC-32G Harrick Plasma Cleaner) for the hydrophilic treatment of the glass substrate. Glass substrates were treated with low power oxygen plasma for 3 min, the contact angle is 10°. Glass substrates were treated with low power oxygen plasma for 15 min, and the contact angle is almost 0o.
3
PDMS Microfluidic Device Fabrication
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Devices were
replicated from the
3D-printed mold in PDMS by first mixing silicone elastomer and curing
agent (Sylgard 184, Dow Corning) at a 10:1 ratio for 5 min at 2000
rpm in a Thinky ARE-310 mixer, followed by a degassing step at 2200
rpm for 5 min. The PDMS mixture was poured on the 3D printed mold,
degassed again in a vacuum desiccator for 20 min, and then cured at
75 °C for 2 h. The cured PDMS was peeled off from the mold, and
1.5 mm diameter reservoirs were punched in the gel and fluid channels.
The devices were bonded to a glass slide after oxygen plasma treatment
for 2 min in a Harrick PDC-32G plasma cleaner and then heated at 75
°C for 36 h prior to use.
replicated from the
3D-printed mold in PDMS by first mixing silicone elastomer and curing
agent (Sylgard 184, Dow Corning) at a 10:1 ratio for 5 min at 2000
rpm in a Thinky ARE-310 mixer, followed by a degassing step at 2200
rpm for 5 min. The PDMS mixture was poured on the 3D printed mold,
degassed again in a vacuum desiccator for 20 min, and then cured at
75 °C for 2 h. The cured PDMS was peeled off from the mold, and
1.5 mm diameter reservoirs were punched in the gel and fluid channels.
The devices were bonded to a glass slide after oxygen plasma treatment
for 2 min in a Harrick PDC-32G plasma cleaner and then heated at 75
°C for 36 h prior to use.
4
Fabrication of Modified Carbon Tape Electrodes
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The fabrication of the modified carbon tape electrodes has been reported previously.25 (link) Briefly, a piece of carbon tape (8 mm long and 7 mm wide) was attached on a piece of the conductive ITO glass (20.0 mm long and 7 mm wide) and then modified with 15 μL 0.025 mg mL−1 multi-wall carbon nanotubes (150 μL, 2 mg mL−1 multi-wall carbon nanotubes and 850 μL water). A piece of transparent tape with a hole was applied on the dried carbon tape to provide effective area for electrochemical detection.25 (link) The modified electrodes were treated under oxygen plasma for 1 min in a PDC-32 G plasma cleaner (Harrick Plasma, Ithaca, NY). Then a piece of circular filter paper was applied on the modified electrodes for detection.25 (link) It needs to emphasize the filter paper could not only store the buffer solution but also provide conductive connection among the electrodes.
5
Cell Adhesion Protocol for Yeast
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Plastic 50 × 9 mm petri dishes (Corning Inc., Corning, NY) were prepared for yeast cell adhesion by plasma cleaning for 3 min with PDC‐32G plasma cleaner (Harrick Plasma, Ithaca, NY). Plates were then incubated for 48 h at 4°C with 0.8 mg/mL CellTak adhesive (Corning Life Sciences, Glendale, AZ) diluted in 0.1‐M NaHCO3. Treated dishes were washed twice with ddH2O to remove excess CellTak and allowed to air dry for 5 min prior to plating cells.
6
Fabrication of 3D and 2D Substrates
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3D CNFs were obtained from the laboratory of M.D.C. as previously reported (52 , 54 , 55 (link)) (see the Supplementary Materials for details). The bulky 3D scaffolds were cut into thin square slices (lateral size, 3 mm × 4 mm; thickness, 250 to 400 μm) and then secured on standard glass coverslips (Kindler) by PDMS (Sylgard 184 silicone elastomer, Dow Corning) cured at 150°C for 15 min. Thereafter, substrates were cleaned under low-pressure air plasma for 5 min (PDC-32G Plasma Cleaner, Harrick Plasma) and ultraviolet (UV)–sterilized for 20 min before use.
PDMS scaffolds are 3D, self-standing, porous microsponges fabricated as described by Bosi et al. (28 (link)). 2D MWCNT supports were obtained as previously described by Fabbro et al. (8 (link)).
PDMS scaffolds are 3D, self-standing, porous microsponges fabricated as described by Bosi et al. (28 (link)). 2D MWCNT supports were obtained as previously described by Fabbro et al. (8 (link)).
7
Microfluidic Cell Growth Monitoring
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Microfluidics experiments were performed as described previously [35 (link)–38 (link)]. Cell growth was imaged within mother machine channels of 25 × 1.4 × 1.26 μm (length × width × height). Within these channels, cells could experience the batch culture medium that diffused through the main flow channels. The microfluidic device consisted of a PDMS flow cell (50 µm/23 µm). The PDMS flow cell was fabricated by mixing the SYLGARD 184 Silicone Elastomer Kit chemicals 10:1 (w/v), pouring the mix on a master waver and hardening it at 80 °C for 1 h. The solid PDMS flow cell was cut out of the master waver and holes were pierced at both ends of each flow channel prior to binding it to a cover glass (Ø 50 mm) by applying the “high” setting for 30 s on the PDC-32G Plasma Cleaner by Harrick Plasma. The flow cell was connected via 40 mm Adtech PTFE tubing (0.3 mm ID × 0.76 mm OD) to a Ismatech 10 K Pump with 40 mm of Ismatech tubing (ID 0.25 mm, OD 0.90 mm) which again was connected via 80 mm Adtech PTFE tubing (0.3 mm ID × 0.76 mm OD) via a 5 mm short Cole-Parmer Tygon microbore tubing (EW-06418-03) (ID 0.762 mm OD 2.286 mm) connector tubing to a Hamilton NDL NO HUB needle (ga21/135 mm/pst 2) that was inserted into the feeding culture. During the whole experiment the pump flow was set to 1.67 µl/min (0.1 ml/h).
8
Preparing Coverslips and Slides for TIRFM
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Coverslips and microscope slides (#1.5; Fisher Scientific) for TIRFM were prepared by washes in acetone, isopropanol, and water followed by sonication for 30 min in isopropanol. Washed glass was then cleaned by plasma cleaning for 3 min using a Harrick PDC-32G plasma cleaner (Harrick Plasma, Ithaca, NY). Cleaned coverslips and microscope slides were immediately passivated by incubation in 1 mg/mL PEG-Si (5000 MW) in 95% ethanol for 18 hr (Winkelman et al., 2014 (link)). Coverslips and slides were then rinsed in ethanol and water, and flow chambers were assembled as described previously (Zimmermann et al., 2016 (link)).
9
Cryo-EM Sample Preparation Protocol
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Four hundred-mesh Cu grids with carbon films (made in-house) were glow discharged for 30 s (Harrick PDC-32G Plasma Cleaner; Harrick Plasma, Ithaca, NY, USA). For sample prep, grids were floated face-down on a 10 μL droplet of sample for 10 min at room temperature, and then rinsed through 3 droplets (20 μL each) of deionized water. Excess liquid was wicked off with a filter paper triangle and grids were incubated for 2 min on 20 μL droplets of 1% uranyl acetate. Excess liquid was again wicked off and grids were left on filter paper to air dry. Grids were imaged at 80 kV in a Hitachi HT7700 TEM and images were collected with an AMT XR16M 16-megapixel digital camera.
10
Fabrication of PDMS-Glass Microfluidic Devices
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Sketches of the PDMS-glass device and dimensions are displayed in the ESI,† Fig. S1. The protocol for the fabrication of the master mould is also included in the ESI.† Polydimethylsiloxane (PDMS) microfluidic devices were fabricated using a 10 : 1 mass ratio of Sylgard 184 silicone elastomer base and curing agent (Dowsil, Midland, MI, United States). These reagents were mixed well using a plastic spatula and degassed using a vacuum pump until visually bubble-free. Approximately 5 g of the mixture was poured onto the master mould and placed at 80 °C for 3 h. Afterwards, the cured PDMS was peeled off from the master mould and unnecessary parts of the PDMS block were cut off. Four inlets/outlets were punched using a 1.5 mm biopsy puncher (Integra LifeSciences, Princeton, NJ, United States). The surface of the device was cleaned using adhesive tape. A No. 1.5 microscopy glass slide (0.16–0.19 mm thick, Biosystems, Muttenz, Switzerland) was cleaned by washing with acetone, isopropanol, and water, then dried using nitrogen gas and a heater at 150 °C. The device and the glass slide were plasma-activated using a PDC-32G plasma cleaner (Harrick Plasma, Ithaca, NY, United States) at less than 0.9 mbar for approximately 1 min and bond together. The glass-bonded device was put on a heater at 150 °C for 5 min and stored at room temperature until use.
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