Mice were anesthetized using intraperitoneal injection of 2% Avertin solution (tribromoethyl alcohol/tertiary amyl alcohol, vol/vol, Sigma-Aldrich), and framed in stereotaxic device (Kopf instruments, CA, USA). Mouse surgery for unilateral implantation of guide cannula was done according to the protocols [5 (link)], targeting the MD (AP: −1.5 mm, ML: 0.3 mm, and DV: −3.3 mm from the brain surface). Gabazine or THIP (from Sigma-Aldrich, St. Louis, MO, USA) were dissolved in normal saline, and a volume of 0.3 μl (30 and 50 μM, respectively) was infused (0.1 μlmin−1) through a 33-gauge injection cannula inserted in the guide cannula. The injector was kept in place for 3 min after the end of the injection. Histologic confirmation of the infusion position was carried out and only the data from properly injected mice were used for statistical analysis.
Avertin solution
Manufactured by Merck Group
Sourced in United States
Avertin solution is a laboratory product manufactured by Merck Group. It is a clear, colorless liquid used as an anesthetic agent for laboratory animals during experimental procedures. The solution contains tribromethanol and amylene hydrate as the active ingredients.
Automatically generated - may contain errors
Lab products found in correlation
Steponeplus rt pcr system, by Thermo Fisher Scientific (2 mentions)
Power sybr green pcr master mix, by Thermo Fisher Scientific (2 mentions)
Rneasy mini kit, by Qiagen (2 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (2 mentions)
Iscript cdna synthesis kit, by Bio-Rad (2 mentions)
Lightcycler 96, by Roche (2 mentions)
Stereotaxic device, by Kopf Instruments (1 mentions)
Gabazine, by Merck Group (1 mentions)
Bleomycin, by Merck Group (1 mentions)
Trametes versicolor, by InvivoGen (1 mentions)
C57bl 6 mice, by Orient Bio (1 mentions)
Rose bengal solution, by Merck Group (1 mentions)
Pericam psi, by Perimed (1 mentions)
2 2 2 tribromoethanol, by Merck Group (1 mentions)
C57bl 6ncrljori, by Orient Bio (1 mentions)
D luciferin, by PerkinElmer (1 mentions)
Living image software, by PerkinElmer (1 mentions)
Ivis lumina 2, by PerkinElmer (1 mentions)
12 protocols using avertin solution
1
Unilateral MD Infusion of GABAA Agonists in Mice
2
Hindlimb Unloading Effects on Rat Bone Marrow
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The experimental design is presented in Figure 5 . Twelve male Wistar rats, 6 months old, weighting 250–300 g, were randomly divided into following experimental groups: Vivarium Control (VC) (n = 6), Hindlimb Unloading (HU) (n = 6). During the experiment, the animals were kept in standard vivarium conditions and administered standard rodent food and water ad libitum (12-h day–night cycle). After 14 days, three animals in each group were selected randomly, sacrificed and BM specimens were prepared for further ex vivo analysis. The other three animals after HU were returned to the animal house. This group was marked as Hindlimb Unloading + Reloading (HU + RL). After 14 days, all animals (VC and HU + RL) were sacrificed and BM samples prepared for further examination. Rats were anesthetized via an intraperitoneal injection of 10% avertin solution at 5 mL/kg of body weight (Sigma-Aldrich Corp., St. Louis, MO, USA) and euthanized via cervical dislocation [38 (link)]. HU was executed according to well-established experimental protocol for small laboratory animals [39 (link),40 ].
All procedures were approved by the Commission on Biomedical Ethics of the Institute of Biomedical Problems (IBMP) (Minutes No. 515 dated 10 June 2019).
All procedures were approved by the Commission on Biomedical Ethics of the Institute of Biomedical Problems (IBMP) (Minutes No. 515 dated 10 June 2019).
3
Trametes versicolor β-glucan Protects against Bleomycin-induced Lung Injury
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All mice were maintained under specific‐pathogen free conditions. Six‐week‐old male C57BL/6 mice were purchased from Orient Bio (Seongnam, Korea) and allowed to adapt for at least 1 week prior to experiments. Mice were randomly assigned to control and treatment groups. Mice were intraperitoneally injected with β‐glucan peptide from the fungus Trametes versicolor (25 mg kg−1; InvivoGen, CA, USA) or PBS. The dose of β‐glucan titrated yielded consistent results. After 7 d, the mice were subjected to bleomycin‐induced lung injury. Under anesthesia with Avertin solution (Sigma), the trachea of each mouse was surgically exposed, followed by intratracheal administration of bleomycin (3 U kg−1; Sigma, B5507) in 40 µL PBS or PBS alone. The mice were sacrificed as indicated, and their BALF, lungs, and bone marrow tissues were further examined.
4
Photothrombosis-Induced Focal Ischemic Stroke
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A focal ischemic stroke was induced at M1FL using the photothrombosis technique as previously described (Takatsuru et al., 2009; Cui et al., 2020; Yao et al., 2020). Briefly, 100 male mice were anesthetized with an intraperitoneal injection of 1.25% avertin solution (80 mg/kg; Sigma-Aldrich, St. Louis, MO, USA). Approximately 10 minutes before surgery, 1.5% Rose Bengal solution (10 μL/g, Sigma-Aldrich), which can induce infarction after laser irradiation, was injected intraperitoneally. The targeted brain region was located in the right M1FL (forelimb region of M1) (coordinates: anterior-posterior (AP), +0.74 mm; medial-lateral (M/L), +1.5 mm) from the superficial) (Paxinos and Franklin, 2013). Irradiation wave length and power were 530 nm and 15mW, respectively, as described previously (Cui et al., 2020; Yao et al., 2020). After 10 minutes of irradiation, the infarcted area was examined using a laser speckle imaging system (PeriCam PSI; Perimed, Stockholm, Sweden).
5
In vivo Lymph Node Imaging in Mice
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For in vivo imaging, C57BL/6 mice were anesthetized with 300 µL of 2.5% avertin solution (2,2,2-tribromoethanol-tertamyl alcohol; Sigma-Aldrich), and the imaging areas were treated with a depilatory cream. aPNM-IRDye800 (50 µg in 50 µL of water) was intradermally injected into the forepaw pad. aPNM-IRDye800 was tracked by using a custom-made whole body optical imaging system at various experimental time points. Near-infrared spectroscopy images (0.5-second exposure) of the axillary lymph nodes were acquired using a 785-nm, 500-mW diode laser as an excitation light source and 835/45-nm band-pass emission filter. All images were processed by using Simple PCI software (Compix Inc., Cranberry Township, PA, USA).
6
Langendorff Cardiac Perfusion in Mice
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Mice were killed by cervical dislocation in accordance with Schedule 1 killing method. Hearts were rapidly excised and Langendorff‐perfused with oxygenated Krebs solution (containing in mM: NaCl 119, NaHCO3 25, sodium pyruvate 1.8, KH2PO4 1.2, KCl 4.7, MgCl2 1.0, CaCl2 1.8, and glucose 10; equilibrated with 95% O2/5% CO2, pH 7.4) at 37°C and at constant rate of 3.5–4 ml/min for 10 min. To prevent formation of blood clots in the coronary circulatory system, animals were treated with heparin (200 units) under non‐recovery terminal general anaesthesia by injection of an overdose of 1.2% Avertin solution (0.5∼0.8 ml i.p. (2,2,2‐tribromoethanol, Sigma‐Aldrich Poole, Dorset, UK) prior to killing and hearts were injected with streptokinase (400 units) via the coronary system after organ excision according to approved procedure by the Home Office.
7
Optical Imaging of Encapsulated Grafts
8
Bleomycin-Induced Pulmonary and Systemic Fibrosis in Mice
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C57BL/6 mice (C57BL/6NCrljOri) were purchased from Orient Bio (Seongnam, Gyeonggi Province, Korea). Del-1−/− mice on a C57BL/6 background were kindly provided by Prof. T. Chavakis (Dresden University, Germany) (24 (link)). Sex- and age-matched mice were used for the experiments. Six- to eight-week-old WT and Del-1−/− mice were anesthetised by intraperitoneal injection of avertin solution (0.5 mL, containing tribromethanol and amylalcohol) (Sigma-Aldrich, St. Louis, MO, USA). Forty microliters of BLM (Sigma-Aldrich, B5507; MB Cell, MB-B4252) were injected into the trachea. Different concentrations (0.7–2 U/kg) of BLM were used for some experiments because BLM activity differed according to the provider and the lot number. At the indicated dpa, the BALF and lung tissues were collected for analysis. To induce systemic fibrosis in WT and Del-1−/− mice, 100 μL of BLM (8 U/kg/dose; Sigma-Aldrich, B5507) were injected subcutaneously on a daily basis for 15 days, as previously described (33 (link)). Skin and lung tissues were collected at 23 dpa after the first BLM injection.
9
In Vivo and Ex Vivo Bioluminescence Imaging
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Bioluminescence imaging (BLI) analysis was performed using the IVIS Lumina II equipped with the Living Image software for data quantification (PerkinElmer, Waltham, MA, USA), as previously described [52 (link)]. For in vivo imaging in particular, mice were sedated with an i.p. injection of Avertin solution (2.2.2-tribromoethanol; Sigma Aldrich) (240 mg/kg) and d -luciferin (PerkinElmer) dissolved in PBS (75 mg/kg body weight) was administered i.p. 10 min before analysis. For ex vivo imaging, animals were euthanized and their organs excised, placed into clear bottom tissue culture dishes, and incubated 10 min before analysis into PBS containing d -luciferin (150 µg/mL) [53 (link)].
10
Murine Thymectomy Procedure
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Thymectomy was performed as previously described58 (link). Briefly, 6-week-old mice were anesthetized by intraperitoneal injection of 2.5% avertin solution (Sigma), and the upper part of the thorax was opened, and a small opening was made in the pleura (1-to 2- cm incision). Then, thymic lobi were carefully removed onto the exterior surface with forceps, and the skin was closed with a puncher.
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