On Days 15 and 16, all rats received Stage II training. Immediately prior to each day of Stage II training, rats received 0.5 μL infusions of either 0.5% (wt/vol) Bupivacaine hydrochloride or 0.9% (wt/vol) saline directed at the MIT. Dummy caps were removed and a 33-gauge microinjection cannula was inserted into the guide cannula, targeting 1 mm below the tip of the guide cannula. The microinjection cannula was attached to a 10 μL Hamilton glass syringe by PE-50 tubing. The glass syringe was operated by an infusion pump (KD Scientific) that infused each solution at 0.25 μL/min. The microinjection cannula was left in place for a further 2 min to permit diffusion into neural tissue. Rats were then placed in the conditioning boxes for Stage II training. During this training, rats received four presentations of CSAB compound co-terminating with the US. The ITI was on average 15 min. Stage II training sessions were 70 min in duration.
Infusion pump
Manufactured by KD Scientific
Sourced in United States
The Infusion pump is a medical device used to deliver fluids, medications, and other solutions at a controlled rate into a patient's body. It is designed to provide accurate and consistent delivery of fluids, ensuring precise dosing and administration.
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Lab products found in correlation
Pe 20, by Braintree Scientific (2 mentions)
Catalog 287571, by Merck Group (2 mentions)
Mp150, by Biopac (2 mentions)
Muscimol, by Merck Group (2 mentions)
Cycloheximide (chx), by Merck Group (2 mentions)
Allopregnanolone, by R&D Systems (1 mentions)
Hydroxypropyl β cyclodextrin, by Merck Group (1 mentions)
26 g needle, by Hamilton Company (1 mentions)
Bone wax, by Johnson & Johnson (1 mentions)
Muscimol, by Bio-Techne (1 mentions)
Bacterial collagenase type 4 s, by Merck Group (1 mentions)
α chloralose, by Merck Group (1 mentions)
β cyclodextrin, by Merck Group (1 mentions)
Ibotenic acid, by Merck Group (1 mentions)
Muscimol mus, by Merck Group (1 mentions)
Compass dataanalysis 4, by Bruker (1 mentions)
Collagenase 7, by Merck Group (1 mentions)
Vt1000, by Leica camera (1 mentions)
Hamilton syringe, by KD Scientific (1 mentions)
Thin walled borosilicate glass, by World Precision Instruments (1 mentions)
37 protocols using infusion pump
1
Bupivacaine Infusion for Conditioning
2
Allopregnanolone Delivery in Rodents
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All drugs were prepared in a 30% (w/v) solution of the complexing agent hydroxypropyl-β-cyclodextrin (VEH; Sigma-Aldrich, St. Louis, MO, USA) in purified water. Allopregnanolone or (3α,5α)-3-hydroxy-pregnan-20-one (ALLO; R&D Systems, Minneapolis, MN, USA) was solubilized in VEH (8 mg/ml). The 5α-reductase inhibitor commonly known as finasteride, (5α,17β)-N-(1,1-dimethylethyl)-3-oxo-4-azaandrost-1-ene-17-carboxamide (FIN; Sigma-Aldrich), was solubilized in VEH (10 mg/ml). The steroid antagonist, 17-phenyl-(3α,5α)-androst-16-en-3-ol (17-PA; R&D Systems) was solubilized in VEH (3.5 mg/ml). Infusions were made with 10-μl Hamilton syringes mounted into an infusion pump (KD Scientific, Hollistan, MA, USA) and connected to internal cannulae (33-gauge, 9 mm with 1-mm projection; Plastics One) with either polyethylene tubing (PE-20, Braintree Scientific, Braintree, MA, USA) for VEH or polytetrafluoroethylene tubing (PTFE; 28-gauge, SAI Infusion Technologies, Lake Villa, IL, USA) for drugs. PTFE tubing was used to minimize drug loss due to nonspecific binding.
3
Flurothyl-Induced Seizure Modeling in Mice
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Flurothyl-induced seizure studies were performed based on prior studies with some modifications (Ozkan et al., 2014 (link); Clement et al., 2012 (link); Dravid et al., 2007 (link)). Briefly, experiments were conducted in a chemical fume hood. Mice were brought to the experimental area at least 1 hr before testing. To elicit seizures, individual mice were placed in a closed 2.4 L Plexiglas chamber and exposed to 99% Bis (2,2,2-triflurothyl) ether (Catalog# 287571, Sigma-Aldrich, St. Louis, MO). The flurothyl compound was infused onto a filter paper pad, suspended at the top of the Plexiglas chamber through a 16 G hypodermic needle and tube connected to a 1 ml BD glass syringe fixed to an infusion pump (KD Scientific, Holliston, MA, USA, Model: 780101) at a rate of 0.25 ml/min. The infusion was terminated after the onset of a hind limb extension that usually resulted in death. Cervical dislocation was performed subsequently to ensure death of the animal. Seizure threshold was measured as latency (s) from the beginning of the flurothyl infusion to the beginning of the first myoclonic jerk.
4
Intracerebral Hemorrhage Mouse Model
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ICH was induced by a double-injection method described previously, 29 (link) with minor modifications. Briefly, 30-μL nonheparinized blood was withdrawn from the angular vein after anesthesia and quickly transferred into a 50-μL syringe with a 26 G needle (Hamilton Company). Then mouse was fixed on a stereotactic frame (RWD Life Science). A 1-mm burr hole was drilled, and blood was infused at the caudate nucleus (2.3 mm lateral to midline, 0.5 mm anterior to bregma, and 3.5mm depth below the surface of the skull). Each ICH model received 30-μL whole blood at a rate of 1 μL/min through the infusion pump (KD Scientific). During the process, needle was paused for 5 minutes before injection, then the first 5 μL was injected to generate a clotting along the needle track, after an additional 5-minute pause, the remaining 25 μL was injected during the following 25 minutes at the same rate of 1 μL/min. After completing the infusion, the needle was held in place for 20 minutes, and then it was slowly withdrawn at a rate of 1 mm/min over the course of 3 movements with 5-minute intervals. Finally, the burr hole was sealed with bone wax (Johnson & Johnson), sterilized and the wound was sutured. After surgery, animals were placed in a cage with free access to food and water. Warming lamps were used to provide thermal support throughout the procedure.
5
Drug Microinfusions in Rat Behavioral Testing
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For drug microinfusions, rats were moved into a room adjacent to the vivarium and placed into 5-gallon white buckets. Dummy cannula internals were removed and a stainless-steel injector (33-gauge, 9 mm; Plastics One) connected to polyethylene tubing was inserted into the guide cannula. Polyethylene tubing was connected to 10-μl Hamilton syringes that were mounted in an infusion pump (Kd Scientific). Muscimol was diluted to a concentration of 0.1 μg/μl in sterile saline. Infusions were made at a rate of 0.3 μl/min for 1 min and the injectors were left in place for 2 min post-infusion to allow for adequate diffusion. Each infusion was verified by the movement of an air bubble that separated the drug or sterile saline from distilled water within the polyethylene tubing. Clean dummy internals were inserted into each guide cannula after infusions. All infusions were made ~5 min prior to behavioral testing. All animals were acclimated to these procedures by performing dummy changes twice before beginning behavioral procedures.
6
Mouse Models of Intracerebral Hemorrhage
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As we previously reported, ICH was induced in mice by injection of autologous blood or bacterial collagenase (Li et al., 2017a (link),b (link); Ren et al., 2018 (link)). First, mice were anesthetized with an intraperitoneal injection of ketamine (100 mg/kg) and xylazine (10 mg/kg). After placing the mice on a stereotactic frame, a 1-mm burr hole was drilled on the right side of the skull (2.3 mm lateral to the midline, 0.5 mm anterior to the bregma). For the collagenase ICH model, 0.0375U bacterial collagenase (Type IV-S, Sigma, St. Louis, MO, USA) dissolved in 0.5 μl saline was infused at the caudate nucleus (3.7 mm depth beneath the skull) through an infusion pump (Kd Scientific Inc., Holliston, MA, USA) at a rate of 0.5 μl/min. In some experiments, we induced the mouse ICH model by infusion of autologous blood using a double-injection method. Whole blood (30 μl) was withdrawn from the angular vein and then infused into the brain as previously described (Sun et al., 2016 (link)). 5 μl of blood was first injected to generate a clot at a depth of 3 mm beneath the hole. Then the needle was moved to a depth of 3.5 mm to inject the remaining 25 μl of blood at the rate of 1 μl/min. After surgery, animals were placed in cages and provided with free access to food and water.
7
Mesolimbic Dopamine Activation in Voles
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Thirty minutes before SPA, 200nL aCSF containing no drug (CSF), 0.4 ng SKF 38393 (SKF), or 4 ng SCH 23390 (SCH), was injected bilaterally with a 1mm projection using 33 gauge internal cannula (PlasticsOne) into the MeA through the guide cannulae. The rate of infusion (200 nl/min) was controlled with an infusion pump (KD Scientific). The subjects were returned to the home cage for 20 min, then placed in the empty testing arena for a 10-min habituation prior to testing for a total 30-min drug activation time. These doses and time period were selected based on previous literature using these drugs in prairie voles (Liu et al., 2011) .
8
GABA Receptor Modulation in Delay Conditioning
9
Precise LHb Inactivation Protocol
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A day before microinjection in experiments 2a and 2b, the injection cannula (Plastics One, Roanoke, VA, USA), which extended 1 mm beyond the guide was inserted into the guide cannula and left in place for 1 min. This was done to control for any initial mechanical damage done by the injector. On a test day, rats were injected with a combination of baclofen and muscimol (Bac/Mus, Sigma) in 0.9% saline, GABA b and a agonists respectively, or vehicle. Both injections used a volume of 0.2 μL (50 ng/0.2 μL baclofen and muscimol) and a 0.15 μL/min infusion rate. This is similar to other LHb inactivation studies that used baclofen and muscimol (Stopper and Floresco, 2014 (link); Mathis et al., 2015 (link)). The injection cannula was connected to a 10 μl syringe (Hamilton) via polyethylene tubing (PE 20) using an infusion pump (KD Scientific).
10
Vascular Reactivity Assessment Protocol
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Intravenous infusions of the selective α1-adrenoceptor agonist phenylephrine (70 μg/mL at 0.4 mL/min/kg), the nitric oxide donor sodium nitroprusside (SNP; 100 μg/mL at 0.8 mL/min/kg), and acetylcholine (10 μg/mL at 1.2 mL/min/kg) was administered using an infusion pump (KD Scientific, Holliston, MA, USA) (Almeida et al., 2015 (link); Duarte et al., 2015 (link)). The infusions of the vasoactive drugs were randomized, and the second treatment was not given before the cardiovascular parameters returned to the control values (the interval between the infusions was approximately 5 min). The infusions lasted for 20–30 s, resulting in the administration of a total dose of 9–14 μg/kg of phenylephrine, 26–40 μg/kg of SNP, and 4–6 μg/kg of acetylcholine.
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