As shown in Figure 1C , the PANDA system was set up by connecting the well plate and a syringe (10 mL, Becton Dickinson, United States) installed in a syringe pump (NE-4000, New Era Pump Systems, United States) through a silicone tubing (1/16 inch inner diameter, EW-95802-02, Cole-Parmer, United States) to generate a constant and steady withdrawal of air from the PANDA chip. The connection between the silicone tubing and syringe was made using a female Luer fitting (FTLL210-9, Nordson MEDICAL, United States). A Luer 3-way valve (Guangzhou JU Plastic Fitting Technology, China) was installed between the PANDA chip and syringe pump, allowing the disconnection of the PANDA chip from the syringe pump after the formation of the hanging drops.
Ne 4000
Manufactured by New Era Pump Systems
Sourced in United States
The NE-4000 is a syringe pump designed for precise fluid dispensing and infusion applications. It features a wide flow rate range, variable speed control, and compatibility with a variety of syringe sizes.
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Lab products found in correlation
9 protocols using ne 4000
1
Hanging Drop Generation using PANDA System
2
Pectin-Based Microparticles Synthesis
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A homemade spray drier was set up using an ultrasonic atomizer with a continuous multiple feeding syringe pump and coupled with a heating column as shown in Figure 1 . An aqueous pectin solution (0.05−0.15 mg/mL, coded as A) with or without theophylline (TH, 0.2 mg/mL) was filled into a syringe, and another aqueous solution containing CHC (0.1, 0.2 mg/mL) or calcium chloride (0.10 mg/mL) as a crosslinking agent (coded as B) was filled into the other syringe. These two solutions, A and B, were then fed into the atomizer cell at a constant flow rate of 0.125 mL/min by a syringe pump (NE-4000, NEW ERA Pump Systems Inc., Farmingdale, NY, USA). The spray drops of the mixed solution would float and be dried in a heated upstream airflow under 1.6 L/min. at controlled temperatures of T1 = 130 °C; T2 = 140 °C; and T3 = 150 °C. The dried particles were collected by a polytetrafluoroethylene membrane (MPT2247, 0.22 µm, ChromTech, Bad Camberg, Germany) and kept in a dry box for further study.
3
Microfluidic Dielectrophoretic Capture and Impedance Sensing
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A custom-made
chip holder based on pogo-pins (Mill-Max Corp.) was employed to electrically
connect our microfluidic device to all measurement equipment. A set
of switches in the holder allowed us to control the signal applied
to each electrode. The flow of spores in the solution within the microfluidic
channel was generated and controlled using a syringe pump (New Era
Pump Systems Inc. NE-4000). During DEP experiments, sinusoidal signals
were applied to the electrodes via the chip holder using a function
generator (Rigol DG822) through a bipolar 10× amplifier (Tabor
Electronics 9250). An oscilloscope (Tektronix TDS 2012B) was also
used to monitor the applied signal. During the process of DEP capture,
our device was placed on the viewing stage of an upright fluorescence
microscope (Amscope FM820TMF143) integrated with a CCD camera (Sony
ICX825ALA) for imaging and video recording. nF-EIS measurements were
performed using a high-precision impedance analyzer (Zurich Instruments
MFIA) controlled by the software LabOne.
chip holder based on pogo-pins (Mill-Max Corp.) was employed to electrically
connect our microfluidic device to all measurement equipment. A set
of switches in the holder allowed us to control the signal applied
to each electrode. The flow of spores in the solution within the microfluidic
channel was generated and controlled using a syringe pump (New Era
Pump Systems Inc. NE-4000). During DEP experiments, sinusoidal signals
were applied to the electrodes via the chip holder using a function
generator (Rigol DG822) through a bipolar 10× amplifier (Tabor
Electronics 9250). An oscilloscope (Tektronix TDS 2012B) was also
used to monitor the applied signal. During the process of DEP capture,
our device was placed on the viewing stage of an upright fluorescence
microscope (Amscope FM820TMF143) integrated with a CCD camera (Sony
ICX825ALA) for imaging and video recording. nF-EIS measurements were
performed using a high-precision impedance analyzer (Zurich Instruments
MFIA) controlled by the software LabOne.
4
Microfluidic Bacterial Mating Assay
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The microfluidic device was fabricated as previously described [51 (link)]. The single channel was placed on the top and in the center of the agarose membrane (20 mm long and 20 mm wide) as Figure 1 a shows. Donor and recipient cells were firstly washed by PBS three times and then diluted to ~108 cells/mL, and the proportion of donor and recipient cells was 1:1 to obtain the initial bacterial solution used for the mating assays. A 5 μL drop of mixed bacterial solution cells were inoculated between the agarose membrane and glass. During the experiments, LB medium was continuously delivered at a speed of 2 μL/min by a syringe pump (NE-4000, NEWERA Pump Systems Inc., Farmingdale, NY, USA) into the channel of the chip for 24 h. The distribution of bacterial growth substrates in the medium was heterogeneous during the experiments, and different thicknesses of biofilms can form between the agarose membrane and glass. At the same time, ARG transfer happened during the biofilm formation, which could be recorded by the microscope. Independently repeated mating assays were performed on the chips three times.
5
Deformability Assessment of d-LON Nanoparticles
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The deformability of d-LON was assessed using a mini-extruder (Avanti Polar Lipids, Alabaster, AL, USA) and a syringe pump (NE-4000, New Era Pump Systems Inc., Farmingdale, NY, USA). A constant pressure of 15.55 mL/h was applied for 2 min to extrude d-LON through 100 nm polycarbonate membranes. The size of the d-LON was measured using DLS, as previously mentioned. The deformability index (DI) of the d-LON was calculated using Equation (1) [26 ]:
where J is the ratio of penetration through the permeable membrane, rv is the average size of the NPs after extrusion, and rp is the pore size of the permeable membrane (100 nm).
where J is the ratio of penetration through the permeable membrane, rv is the average size of the NPs after extrusion, and rp is the pore size of the permeable membrane (100 nm).
6
Microfluidic Antimicrobial Susceptibility
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On-chip inhibition tests were
conducted by delivering “source” and “sink”
solutions through parallel channels in the chip, Figure1 c. Solutions were continuously delivered at a speed of 0.33
mm/s by a syringe pump (NE-4000, Newera Pump Systems Inc.). The “source”
solution contained medium plus either 5 mg/L (E. coli) or 15 mg/L (N. europaea) amoxicillin, and the
“sink” solution contained medium alone. Shortly after
delivery of the two solutions, a steady concentration gradient was
established via diffusion through the agarose membrane to the bacterial
monolayer underneath. To visualize the time course and profile of
the gradient formed, 30 μM fluorescein (MW = 332.31, Sigma-Aldrich)
solution was used in a parallel set of (bacteria free) experiments.
All experiments were performed at room temperature (21–22 °C),
in line with a habitat temperature commonly found for bacteria in
the environment. At least three replicate experiments were conducted
for each of the conditions reported below.
conducted by delivering “source” and “sink”
solutions through parallel channels in the chip, Figure
mm/s by a syringe pump (NE-4000, Newera Pump Systems Inc.). The “source”
solution contained medium plus either 5 mg/L (E. coli) or 15 mg/L (N. europaea) amoxicillin, and the
“sink” solution contained medium alone. Shortly after
delivery of the two solutions, a steady concentration gradient was
established via diffusion through the agarose membrane to the bacterial
monolayer underneath. To visualize the time course and profile of
the gradient formed, 30 μM fluorescein (MW = 332.31, Sigma-Aldrich)
solution was used in a parallel set of (bacteria free) experiments.
All experiments were performed at room temperature (21–22 °C),
in line with a habitat temperature commonly found for bacteria in
the environment. At least three replicate experiments were conducted
for each of the conditions reported below.
7
Multifunctional Microfluidic Cellular Culture Device
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Neuronal cells were cultured in the three middle compartments of the device containing the recording electrodes. Otherwise, endothelial and epithelial cells were cultured in the compartments adjacent to those with the neurons as shown in Fig. 1B.
After seeding, cells were incubated under static conditions for 1.5-3 h to ensure cell attachment.
Then, the cells were perfused with their respective culture media at a rate of 0.1 mL min -1 for 3 days.
For the case of the neuronal cells, the medium was only flowed through the central compartment to simplify the fluidic connections. Culture medium was injected using a syringe pump (NE-4000, New Era Pump Systems, Inc., US) in infusion mode. Polytetrafluoroethylene (PTFE) tubes (0.5 mm internal diameter (ID) and 1 mm outer diameter (OD)) were cut and inserted into the inlets and outlets of the device. Inlet tubes were connected to syringes containing culture media, and the outlet tubes were connected to reservoirs (silicone tube of 0.8 mm ID and 2.4 mm OD). To ensure proper equilibration of the medium with the incubator, reservoirs were vented and placed near the outlets.
After seeding, cells were incubated under static conditions for 1.5-3 h to ensure cell attachment.
Then, the cells were perfused with their respective culture media at a rate of 0.1 mL min -1 for 3 days.
For the case of the neuronal cells, the medium was only flowed through the central compartment to simplify the fluidic connections. Culture medium was injected using a syringe pump (NE-4000, New Era Pump Systems, Inc., US) in infusion mode. Polytetrafluoroethylene (PTFE) tubes (0.5 mm internal diameter (ID) and 1 mm outer diameter (OD)) were cut and inserted into the inlets and outlets of the device. Inlet tubes were connected to syringes containing culture media, and the outlet tubes were connected to reservoirs (silicone tube of 0.8 mm ID and 2.4 mm OD). To ensure proper equilibration of the medium with the incubator, reservoirs were vented and placed near the outlets.
8
Hemodilution and Hemodynamic Monitoring
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Forty percent of BV was exchanged with the test solution in animals, lowering the systemic Hct by 45%. Total BV was estimated as 7% of body weight. Test solutions were infused into the jugular vein catheter at a rate of 100 μl/min and blood was simultaneously withdrawn at the same rate from the left carotid artery using a syringe pump (NE-4000, New Era Pump Systems, USA). After hemodilution, animals were followed over 1 h. MAP and HR were continuously monitored and blood was collected at the end of each experiment for measurements of viscosity. Figure 1 illustrates the experimental protocol.
9
Microfluidic Chip Preparation and Functionality Testing
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Before experiments, the microfluidic chips and cell reservoir were washed with soap and rinsed with deionized water and 70% ethanol. The chips were dried with pressurized air. Then, a piece of scotch tape was used to clean away any remaining dust or particles. For testing the functionality of microfluidics before cell culture experiments, water was pumped with a microfluidic syringe pump (NE-4000, New Era Pump Systems Inc, Farmingdale, NY, USA) with a flowrate of 0.1 mL/h or 1.6 µL/min into the microfluidic channels. If any leaks were detected during this test, the chip was discarded.
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