The animals were anesthetized with an intraperitoneal injection of ketamine 100 mg/kg (Fort Dodge Animal Health, Fort Dodge, Iowa) and Xylazine 6 mg/kg (Phoenix Scientific, Inc., St. Joseph, MO). The glottis was directly visualized and intubated with a 18 gauge Angiocatheter (forced oscillation measurements)(BD Insyte, Sandy, Utah) and transferred to a FlexiVent rodent ventilator (Scireq, Montreal, QC Canada) as previously described [12 (link)]. Ventilator rates of 200/minute, tidal volume of10 ml/kg with positive end-expiratory pressures of 3 cmH2O and a pressure limit of 30 cmH2O were used for the pneumonectomy procedure.
Flexivent rodent ventilator
Manufactured by SCIREQ
Sourced in Canada
The FlexiVent is a rodent ventilator designed for use in laboratory research. The device is capable of measuring respiratory mechanics in small animals such as mice and rats. It provides precise control over ventilation parameters and enables the assessment of lung function.
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Lab products found in correlation
9 protocols using flexivent rodent ventilator
1
Rodent Anesthesia and Ventilation for Pneumonectomy
2
Pulmonary Function Evaluation in Rodents
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Pulmonary function tests were performed in all animals before micro-CT acquisition. To this end, animals were first anesthetized with an intraperitoneal injection of 90 mg/kg ketamine (Imalgene®, Merial, France) and 10 mg/kg xylazine (Rompun®, Bayer AG, Germany). Anesthetized animals were intratracheally cannulated and connected to a Flexivent rodent ventilator (Scireq, Montreal, Canada) set at a rate of 200 breaths/min and a tidal volume of 10 ml/kg. Animals were kept breathing isoflurane at 2% concentration until completely relaxed. Lung resistance (R, measured in cmH2O.s/mL), compliance (C, measured in mL/cmH2O) and elastance (E, measured in cmH2O.s/mL) parameters were measured by using a single-frequency-forced oscillation, fitting the measured data to a single compartment model of the lung. All measurements were repeated at least five times.
3
Micro-CT Imaging of Mouse Lung Volumes
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Thorax tomography was performed on mice anesthetized by intraperitoneal injection of ketamine and xylazine. Then, an intratracheal cannula was placed and connected to a flexiVent rodent ventilator (Scireq) set at a rate of 200 breaths/minute and a tidal volume of 10 mL/kg. Animals were maintained on 2% isoflurane inhalation until completely relaxed.
Lung 3D tomographic images were acquired using an X-ray micro-CT system (Quantum-GX, Perkin Elmer) with the following parameters: X-ray source voltage of 90 kVp, current of 88 μA, high-speed scan protocol for a total acquisition time of 14 minutes, and gantry rotation of 360°. Breathing artifacts were denoised using respiratory gating in each acquisition. The three-dimensional tomographic images containing the whole lung had a total of 512 slices with an isotropic voxel size of 72 μm and a resolution of 512 × 512 pixels per slice. Analysis of the lung volume in each sample was carried out using Fiji/ImageJ (RRID:SCR_002285), open-source Java-based image processing software. In brief, lung images were segmented by applying a fixed threshold, and total lung volume was measured over the obtained mask (mm3). Quantified values were normalized to those of normal, healthy lung parenchyma.
Lung 3D tomographic images were acquired using an X-ray micro-CT system (Quantum-GX, Perkin Elmer) with the following parameters: X-ray source voltage of 90 kVp, current of 88 μA, high-speed scan protocol for a total acquisition time of 14 minutes, and gantry rotation of 360°. Breathing artifacts were denoised using respiratory gating in each acquisition. The three-dimensional tomographic images containing the whole lung had a total of 512 slices with an isotropic voxel size of 72 μm and a resolution of 512 × 512 pixels per slice. Analysis of the lung volume in each sample was carried out using Fiji/ImageJ (RRID:SCR_002285), open-source Java-based image processing software. In brief, lung images were segmented by applying a fixed threshold, and total lung volume was measured over the obtained mask (mm3). Quantified values were normalized to those of normal, healthy lung parenchyma.
4
Lung Tissue Harvesting and Preparation
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After the induction of anesthesia with intraperitoneal injection of ketamine 100 mg/kg (Fort Dodge Animal Health, Fort Dodge, IA, USA) and xylazine 10 mg/kg (Phoenix Scientific, St. Joseph, MO, USA), the animal was endotracheally intubated with an 18G angiocatheter (Becton Dickinson, Franklin Lakes, NJ, USA) and ventilated with the Flexivent rodent ventilator (SciReq, Montreal, Quebec, CA). The animal was euthanized by exsanguination through the inferior vena cava. A median sternotomy facilitated exposure of the anterior mediastinum. In sequence, the left atrium, right ventricle and inferior vena cava were incised. A 22G olive-tipped cannula was inserted through the right ventricle into the pulmonary artery and the lungs were flushed with 20 cc of phosphate-buffered saline at 23oC. A cervical tracheotomy was performed and the orotracheal tube replaced with a second 18G angiocatheter positioned in the distal trachea and secured with a silk tie. The lungs were inflated to 70% total lung capacity (TLC, based on the average of the volumes previously recorded on the Flexivent). During 70% TLC static inflation, the pulmonary artery was flushed first with 20cc of phosphate-buffered saline at 23oC prior preparation for precision-cut lung slices.
5
Quantitative Lung Imaging in Mice
6
Anesthesia and Intubation in Mice
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Twenty-one days after BLM or saline treatment, mice were anesthetized with 50 mg/kg pentobarbital i.p. (Sigma, USA), and tracheostomy was performed. Animals were intubated with a 14-gauge cannula and connected to a Flexivent rodent ventilator (SCIREQ Inc., Canada).
7
Thorax Tomography in Lung Cancer Mouse Model
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Thorax tomography was performed on day 23 after K7M2 cell inoculation. Mice were anesthetized by intraperitoneal injection of ketamine and xylazine and maintained on 2% inhaled isoflurane until they were completely relaxed. Mouse lung inhalation was controlled by placing an intratracheal cannula in mice connected to a flexiVent rodent ventilator (SCIREQ, Montreal, Canada), which was set at a rate of 200 breaths/min and a tidal volume of 10 mL/kg 3D.
Three-dimensional lung tomographic images were acquired using an X-ray microCT system (Quantum-GX; PerkinElmer, Waltham, MA, USA) with the following parameters: X-ray source voltage of 90 kVp, current of 88 μA, high-speed scan protocol for a total acquisition time of 4 min, and gantry rotation of 360°. Breathing artifacts were denoised using respiratory gating in each acquisition. Three-dimensional tomographic images containing the whole lung included a total of 512 slices with an isotropic voxel size of 72 μm and a resolution of 512 × 512 pixels per slice. Analysis of the lung volume in each sample was carried out using Fiji/ImageJ, which is open-source Java-based image-processing software. In brief, lung images were segmented by applying a fixed threshold, and total lung volume was measured over the obtained mask (mm3).
Three-dimensional lung tomographic images were acquired using an X-ray microCT system (Quantum-GX; PerkinElmer, Waltham, MA, USA) with the following parameters: X-ray source voltage of 90 kVp, current of 88 μA, high-speed scan protocol for a total acquisition time of 4 min, and gantry rotation of 360°. Breathing artifacts were denoised using respiratory gating in each acquisition. Three-dimensional tomographic images containing the whole lung included a total of 512 slices with an isotropic voxel size of 72 μm and a resolution of 512 × 512 pixels per slice. Analysis of the lung volume in each sample was carried out using Fiji/ImageJ, which is open-source Java-based image-processing software. In brief, lung images were segmented by applying a fixed threshold, and total lung volume was measured over the obtained mask (mm3).
8
Anesthesia and Mechanical Ventilation in Rodents
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The animals were anesthetized with an intraperitoneal injection of Ketamine 100 mg/kg (Fort Dodge Animal Health, Fort Dodge, Iowa) and Xylazine 10 mg/kg (Phoenix Scientific, Inc., St. Joseph, MO). Prior to mechanical ventilation, the glottis was directly visualized and intubated with a standard 18 gauge Angiocatheter to minimize tracheal air leak (BD Insyte, Sandy, Utah). After intubation, the animal was transferred to a FlexiVent rodent ventilator (Scireq, Montreal, QC Canada) as previously described (Gibney et al. 2011 ).
9
Quantification of Murine Lung Volume via Micro-CT
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Thorax tomography scans were performed on mice anesthetized with an i.p. injection of ketamine and xylazine. Afterward, the mice were intratracheally cannulated and connected to a flexiVent rodent ventilator (Scireq, Montreal, QC, Canada) set at a rate of 200 breaths/min and a tidal volume of 10 mL/kg. Animals were kept breathing 2% isoflurane until completely relaxed.
Tomographic 3D images of the lungs were acquired using X-ray micro-CT (Quantum-GX; Perkin Elmer, Waltham, MA, USA) with the following parameters: 90 kVp X-ray source voltage, 88 μA current, and the high-speed scan protocol for a total acquisition time of 14 min and a gantry rotation of 360 degrees. Breathing-derived artifacts were de-noised using respiratory gating in each acquisition. Tomographic 3D images containing a whole lung had a total of 512 slices with an isotropic 72-μm voxel size and a resolution of 512 × 512 pixels per slice. The analysis of lung volume in each sample was carried out using Fiji/ImageJ, an open-source Java-based image-processing software. Briefly, lung images were segmented by applying a fixed threshold, and total lung volume was measured over the obtained mask (cubic millimeters). Quantifications were normalized to the results for normal, healthy lung parenchyma.
Tomographic 3D images of the lungs were acquired using X-ray micro-CT (Quantum-GX; Perkin Elmer, Waltham, MA, USA) with the following parameters: 90 kVp X-ray source voltage, 88 μA current, and the high-speed scan protocol for a total acquisition time of 14 min and a gantry rotation of 360 degrees. Breathing-derived artifacts were de-noised using respiratory gating in each acquisition. Tomographic 3D images containing a whole lung had a total of 512 slices with an isotropic 72-μm voxel size and a resolution of 512 × 512 pixels per slice. The analysis of lung volume in each sample was carried out using Fiji/ImageJ, an open-source Java-based image-processing software. Briefly, lung images were segmented by applying a fixed threshold, and total lung volume was measured over the obtained mask (cubic millimeters). Quantifications were normalized to the results for normal, healthy lung parenchyma.
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