Following baseline PIT testing for subjects treated with muscimol, rats were anesthetized with a mixture of ketamine (100 mg/kg: Vedco) and xylazine (10 mg/kg) via intraperitoneal (IP) injection (0.1% bodyweight). Subjects were placed in a Kopf (David Kopf Instruments; Tujunga, CA) stereotaxic instrument, and an incision was made over the midsagittal line to reveal bregma and lambda on the surface of the skull. Stainless steel (22 gauge) guide cannula (Plastics One, Roanake VA) were fixed in place with jeweler’s screws and dental cement 1.5 mm above CeA at -2.5 mm posterior, 4 mm lateral to the midline, and 6 mm ventral from the surface of the skull. Subjects recovered in the home cage for one week following surgery and then underwent further PIT testing. Prior to testing, muscimol (0.3 μl of 1 ng/nl solution) or deionized water was infused through (28 gauge) injectors extending 1.5 mm beyond the guides bilaterally at a rate of 0.15 μl/min with subjects connected to the infusion lines for an additional min for dispersal. This was accomplished using 10 μl Hamilton syringes (Model 701N) controlled by a Harvard Apparatus pump (PHD 22/2000) via polyethylene tubing connected to injectors extending 1.5 mm beyond the tip of their guide cannula. Subjects were tested 15–20 min after treatment.
Phd 22 2000
Manufactured by Harvard Apparatus
Sourced in United States
The PHD 22/2000 is a versatile syringe pump designed for a wide range of laboratory applications. It features a flow rate range of 0.0001 to 2000 mL/min and can accommodate syringes from 0.5 μL to 140 mL. The pump offers programmable flow, volume, and time functions, as well as multiple operational modes to suit various experimental needs.
Automatically generated - may contain errors
Lab products found in correlation
Gelatin, by Merck Group (2 mentions)
Ketamine, by Vedco (1 mentions)
Sylgard 184, by Dow (1 mentions)
Pe20 tubing, by Thermo Fisher Scientific (1 mentions)
Way100635, by Bio-Techne (1 mentions)
Mdl100907, by Bio-Techne (1 mentions)
Focus 220, by Siemens (1 mentions)
Quanta 600f, by Thermo Fisher Scientific (1 mentions)
Chcl3, by Merck Group (1 mentions)
Vhx 500fe, by Keyence (1 mentions)
18 protocols using phd 22 2000
1
Intra-CeA Muscimol Infusion for Pavlovian Approach
2
Optimized Microfluidic Biosensing Setup
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The experimental setup, as mentioned in previous paragraphs, was improved for biosensing experiments. The samples were placed in a flow chamber, which contained a micro fluidic polydimethylsiloxane (PDMS, Dow Corning “Sylgard 184”) channel (2 × 15 × 0.5 mm). The fluid inside the microfluidic channel was driven by a syringe pump (Harvard Apparatus PHD 22/2000, Holliston, MA, USA), allowing for well-defined flow rates of 100 µL/min. In addition to the backgate, a reference electrode (Ag/AgCl, Microelectrodes Inc., USA) was introduced to the tubing system. Gate voltages were applied to the backgate and reference electrode of the system to minimize electrical noises. The experimental setup, including electrical circuit, is schematized in Figure S1B .
The applied gate voltage was reduced to a range from −1 to 1V when operating in liquid surroundings. For real-time measurements, a constant gate sweeping mode was realized, where VG was swept continuously and ISD was recorded.
The applied gate voltage was reduced to a range from −1 to 1V when operating in liquid surroundings. For real-time measurements, a constant gate sweeping mode was realized, where VG was swept continuously and ISD was recorded.
3
Microfluidic Magnetic Particle Manipulation
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The sample solution was injected into the microfluidic device using 1 mL syringes and tygon tubing. Syringe pumps (PHD22/2000, Harvard Apparatus, USA) were used to maintain a constant flow rate. A non-uniform magnetic field was generated using two neodymium magnets placed in the microfluidic device. Particle trajectories were observed using an inverted fluorescence microscope (Olympus IX73, Japan). To gain comparable fluorescent data, the exposure time of the camera was fixed to 1 second.
4
Fabrication of 3D PCL Nanofibrous Scaffold
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As shown in the schematic diagram in Additional file 1 (Figure S1), the 3D PCL nanofibrous scaffold was fabricated by electrospinning. Briefly, pellets of PCL with an average molecular weight of 80 kDa (Aldrich, St. Louis, MO, USA) were dissolved in a chloroform/methanol (3:1, v/v) solvent mixture at room temperature (25 ± 1 °C) to obtain a 20 wt % solution. Then, the PCL solution was placed in a 10-ml plastic syringe connected to a blunt stainless steel needle with an inner tip diameter of 0.5 mm. A syringe pump (PHD22/2000, Harvard Apparatus, Holliston, MA, USA) was used to control the flow rate at 2 ml/h. A 20-kV high-voltage power supply and a distance of approximately 15 cm were maintained between the needle and the grounded flat aluminum plate collector (size: 15 × 15 cm2) throughout the electrospinning process. Then, the PCL fibrous scaffolds were sterilized under ultraviolet (UV) light for 1 h and washed three times with 70 % ethanol for 5 min each. Finally, the scaffold was pre-coated with gelatin (Sigma-Aldrich, St. Louis, MO, USA) by immersion in a sterilized 0.1 % (1 g/l) gelatin solution overnight.
5
Intracerebroventricular Drug Infusion in Mice
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For experiments requiring intracerebroventricular (i.c.v.) drug infusion, pre-canulated (third ventricle) C57BL/6J mice were received from The Jackson Laboratory. Animals had internal guide cannula (2.5 mm long; Plastics One, USA) mounted in the 3rd ventricle (standard coordinates—ML: +1.0, RC: −0.4, DV: 2.0 mm) and protected by a dummy cap until the experiment. For more information on the cannulation procedure, animal care, and use, see the link: https://www.jax.org/-/media/jaxweb/files/jax-mice-and-services/brain-cannulation-information-care-use.pdf?la=en&hash=FD75F73AB0CD7A47808C78D0FC405AB3AF123F3B (accessed on 14 June 2021).
Conscious freely moving mice were infused i.c.v. with rimonabant (2 µg), JD5037 (1 µg), or their solvents. Drugs were applied in a volume of 1 μL over the course of 2 min via 33 g internal injector (P1 Technologies, Roanoke, VA, USA) connected with the 2 µL precision glass Hamilton syringe (USA) by a PE-20 tubing (Fisher Scientific, Hampton, NH, USA). The infusion rate and volume were controlled through the use of the syringe pump (model PHD 22/2000, Harvard Apparatus, Cambridge, MA, USA). Following 2 min infusion, injectors were left in place for additional 3 min to allow a passive diffusion of drugs into the tissue.
Conscious freely moving mice were infused i.c.v. with rimonabant (2 µg), JD5037 (1 µg), or their solvents. Drugs were applied in a volume of 1 μL over the course of 2 min via 33 g internal injector (P1 Technologies, Roanoke, VA, USA) connected with the 2 µL precision glass Hamilton syringe (USA) by a PE-20 tubing (Fisher Scientific, Hampton, NH, USA). The infusion rate and volume were controlled through the use of the syringe pump (model PHD 22/2000, Harvard Apparatus, Cambridge, MA, USA). Following 2 min infusion, injectors were left in place for additional 3 min to allow a passive diffusion of drugs into the tissue.
6
Optogenetic Modulation of Fear Conditioning
7
In situ Intestinal Perfusion in Mice
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In situ perfusion experiments was performed with mice and a perfusion pump (PHD22/2000, Harvard Apparatus) as previously described 48 (link). In brief, Bmal1fl/fl and Bmal1iKO mice (male, n = 5) were fasted overnight before experimentation. Mice were anaesthetized with pentobarbital sodium (60 mg/kg) by intraperitoneal injection. A midline longitudinal incision was made to expose the abdomen, and then a segment of jejunum (10 cm) was separated. The intestinal segment was cannulated with a silicone tube at the proximal and distal end of the segment. After removal of the intestinal content and preincubation of the segment with transport medium (HBSS, pH 7.4) containing 20 μM MTX for 30 min, the effluents were collected every 30 min for 120 min. At the end of experiment, the lengths of intestinal segments were measured. MTX concentrations in perfusate samples were determined by LC-MS/MS analysis.
8
Fabrication of Electrospun Polymer Membranes
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Fabrication of RES was carried out by using 1 mL syringes with 20 gauge 1″ syringe tips (8001213, Fisnar, Germantown, WI, USA) loaded with ~1.1 mL of the polymer solution. The syringe was loaded into a syringe pump (PHD 22/2000, Harvard Apparatus, Holliston, MA, USA) and the syringe cap was connected to a voltage supply. The static collector was covered with silicon-coated paper and placed at a distance of 12 cm from the needle tip. Flow rate and voltage are detailed for every solution using their code as a reference in Table 2 . Electrospun membranes were stored at 4 °C in sealable bags.
9
Fabrication of Sintered Microsphere Scaffolds
10
In Vivo PET Imaging of Neuroreceptor
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Prior to each scan, animals were sedated with a combination of alfaxalone (2 mg/kg), midazolam (0.3 mg/kg), and dexmedetomidine (0.01 mg/kg) and maintained in an anesthetized state with 1.5–2.5% isoflurane. Heart rate, blood pressure, respiration rate, and oxygen saturation were monitored continuously. A catheter was placed in the radial artery for blood sampling. All PET scans were performed on a Focus 220 scanner (Siemens Medical Solutions, Knoxville, TN). An 8.5-min transmission scan was collected first for attenuation correction, followed by bolus injection of radiotracer in 10 mL over 3 min by an infusion pump (PHD 22/2000; Harvard Apparatus). Dynamic PET data were acquired in list mode for 120 min starting concurrent with the beginning of the injection, then binned into frames of increasing durations (6 × 30 s, 3 × 1 min, 2 × 2 min, 22 × 5 min). For pre-blocking scans, PF-367 (0.03, 0.01, or 0.25 mg/kg) was administered as a 5-min IV slow bolus beginning 15 min prior to radiotracer injection, then a 120-min scan was acquired as described.
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