The skulls of P90 iZEG-Dlx2 and Wnt1Cre::iZEG-Dlx2 mice were collected and fixed with 4% paraformaldehyde (PFA). Subsequently, micro-CT data were collected using an eXplore Locus MicroCT scanner (GE Healthcare Life Sciences, Milwaukee, WI, USA), using 0.01-mm-thick slices. The 3D reconstructions of the skulls and bone mineral density calculations were completed using GE MicroView software version 2.2 (GE Healthcare Life Sciences).
Explore locus microct scanner
Manufactured by GE Healthcare
Sourced in United States, United Kingdom
The EXplore Locus MicroCT scanner is a laboratory-based X-ray computed tomography (microCT) system designed for high-resolution 3D imaging of small samples. The scanner utilizes a cone-beam X-ray source and a high-resolution digital detector to capture detailed volumetric data of the internal structure of various specimens.
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12 protocols using explore locus microct scanner
1
Micro-CT Analysis of Mouse Skulls
2
Micro-CT Analysis of Adipose Tissue
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Mice were anesthetized with a continuous flow of 3% isoflurane/oxygen mixture and positioned prone with both legs at right angles. The region spanning the entire torso to the distal tibia of each mouse was scanned with Explore Locus micro-CT scanner (GE Healthcare) without contrast agents. Micro-CT projections of the animals were acquired using 80 kV, 450 μA current, with 100 ms acquisition time. Four hundred projections were acquired over 360°. The resolution of the acquired images was 93 μm. The reconstructed 3D images were visualized and analyzed using MicroView analysis software (GE Healthcare).
The amount of adipose tissue was determined as described by Luu et al.[40 (link)]. Briefly, the gray-scale histogram of the reconstructed images presents a peak that indicates the presence of fat. A low and a high gray-scale threshold corresponding to that peak were chosen. The fat was quantified as the sum of the volumes of all the voxels with a gray-scale value ranging between the low and the high thresholds. The analysis was done on the abdominal region (between the proximal end of L1 and the distal end of S1).
The amount of adipose tissue was determined as described by Luu et al.[40 (link)]. Briefly, the gray-scale histogram of the reconstructed images presents a peak that indicates the presence of fat. A low and a high gray-scale threshold corresponding to that peak were chosen. The fat was quantified as the sum of the volumes of all the voxels with a gray-scale value ranging between the low and the high thresholds. The analysis was done on the abdominal region (between the proximal end of L1 and the distal end of S1).
3
Quantifying Adipose Tissue in Mice via Micro-CT Scanning
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In the second set of experiments, mice were anesthetized with a continuous flow of 3% isoflurane/oxygen mixture and positioned prone with both legs at right angles. The region spanning the entire torso to the distal tibia of each mouse was scanned with an Explore Locus micro-CT scanner (GE Healthcare, Chicago, IL, USA) without contrast agents. Four hundred micro-CT projections of the animals were acquired over 360° using 80 kV, 450 μA current and 100 ms of acquisition time with a resolution of 93 μm. The reconstructed 3D images were visualized and analyzed using MicroView analysis software (GE Healthcare, Chicago, IL, USA). The amount of adipose tissue was determined as in [89 (link)]. The gray-scale histogram of the reconstructed images presents a peak that indicates the presence of fat. One low and one high gray-scale threshold corresponding to that peak were chosen. The fat was quantified as the sum of the volumes of all the voxels with a gray-scale value between the low and the high thresholds. The analysis was done on the abdominal region (between the proximal end of lumbar vertebrae L1 and L5).
4
Lung Volume Measurement via Micro-CT
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Chest CT imaging and measurements of lung volumes were performed as previously described [58 (link)–61 (link)]. Mice anesthetized with ketamine/xylazine were maintained under anesthesia using an isoflurane/oxygen mixture to minimize spontaneous breaths throughout the procedure. Images were obtained using the Explore Locus Micro-CT Scanner (GE Healthcare, Waukesha, WI). CT scans were performed at a resolution of 93 mm. Microview software version 2.2 (http://microview.sourceforge.net ) was used to analyze lung volumes and to render three-dimensional images. Lung volumes were calculated from renditions collected at full inspiration.
5
Assessing Lung Function and Imaging in Mice
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Mice were intraperitoneally injected with ketamine/xylazine (100 mg/kg Ketamine, 20 mg/kg Xylazine). Once the correct anesthetic plane was achieved, the mice were intubated with a sterile, 20-gauge intravenous cannula through the vocal cords into the trachea. Following intubation, anesthesia was maintained using isoflurane.
Elastance (Ers), compliance (Crs), and total lung resistance (Rrs) was assessed for each mouse through the snapshot perturbation method, as previously described36 (link). Measurements were performed in triplicates for each animal, using the FlexiVent system (SCIREQ, Tempe, AZ), with a tidal volume of 30 mL/kg at a frequency of 150 breaths/min against 2–3 cm H2O positive end-expiratory pressure.
After PF testing, the mice were subjected to CT scans for the measurements of lung volume, using the Explore Locus Micro-CT Scanner (General Electric, GE Healthcare, Wauwatosa, WI). CT scans were performed during full inspiration and at a resolution of 93 μm. Lung volumes were calculated from lung renditions collected at full inspiration. Microview software 2.2 (http://microview.sourceforge.net ) was used to analyze lung volumes and render three-dimensional images.
Elastance (Ers), compliance (Crs), and total lung resistance (Rrs) was assessed for each mouse through the snapshot perturbation method, as previously described36 (link). Measurements were performed in triplicates for each animal, using the FlexiVent system (SCIREQ, Tempe, AZ), with a tidal volume of 30 mL/kg at a frequency of 150 breaths/min against 2–3 cm H2O positive end-expiratory pressure.
After PF testing, the mice were subjected to CT scans for the measurements of lung volume, using the Explore Locus Micro-CT Scanner (General Electric, GE Healthcare, Wauwatosa, WI). CT scans were performed during full inspiration and at a resolution of 93 μm. Lung volumes were calculated from lung renditions collected at full inspiration. Microview software 2.2 (
6
Visualizing Lung Tumors via microCT
7
Hybrid FMT-XCT Imaging System for Samples
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A fluorescence molecular tomography-X-ray computed tomography (FMT-XCT) hybrid imaging system was used to image the samples. The FMT-XCT system has been described in details in [58] (link). Briefly, the system operates in 360 ° trans-illumination geometry, where the sample was illuminated by a 750 nm laser at 18 equally spaced gantry locations, where images were acquired at both excitation and fluorescence wavelengths of DiR (∼750 and ∼830 nm, respectively) using a CCD cooled at −80 °C. The optical part of the FMT-XCT imaging lasted around 50 min per sample. CT imaging was performed afterwards using the X-ray sub-system of the FMT-XCT, consisting of an eXplore Locus micro-CT scanner (GE Healthcare, U.K.) covering a scan field of ∼40 mm in the axial direction. The CT acquisition lasted ∼20 min per sample. The FMT-XCT system was modeled using a finite-element approach, accounting for light propagation. Anatomical information was used in conjunction with optical data for improved accuracy using a regularized linear least squares approach [46] , [58] (link).
8
Micro-CT Imaging of Mouse Skulls
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Mice were anesthetized intraperitoneally with pentobarbital sodium before micro-CT scans were performed. The slice thickness for the micro-CT scans was 40 μm using an eXplore Locus micro-CT scanner (GE Healthcare, Milwaukee, WI, United States). Three-dimensional (3D) reconstruction of the skulls was performed using GE MicroView software (GE Healthcare, Milwaukee, WI, United States). The bone volume was measured using GeomagicStudio software (Geomagic, NC, United States).
9
Micro-CT Analysis of Mouse Skulls
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The skulls/mandibles of the P90 control and Wnt1Cre::iZEG-Dlx2 mice were fixed with 4% paraformaldehyde at room temperature for 24 h. Next, the tissues were scanned using an eXplore Locus MicroCT scanner (GE Healthcare Life Sciences, Milwaukee, WI, USA). The slice thickness used for micro CT scans was 10 µm. Reconstruction of 3D skulls and bone mineral density (BMD) calculations were performed using GE MicroView software version 2.2 (GE Healthcare Life Sciences).
10
In Vivo Micro-CT Quantification of Body Fat
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For in vivo micro-CT scanning, mice were anesthetized with 1% isoflurane inhalation and a whole-body scan was performed at a resolution of 93 μm (80 kV, 450 μA and 100 ms integration time) using an eXplore Locus Micro-CT scanner (GE Healthcare, Little Chalfont, Buckinghamshire, UK). The micro-CT scanner was calibrated according to protocols provided by the manufacturer. Each scan took approximately 10 min and the mice showed no sign of discomfort during the procedure. The images obtained were analyzed using Microview software (GE Healthcare). Using the Microview software, WAT quantification was assessed from the base of the lungs to the distal side of the hip joint. For determination of total AT volume, histogram analysis of CT images showed that voxels with CT values between -380 and -120 Hounsfield units (H.U) are defined as AT. The abdominal muscular wall was used as the differentiation line to separate sWAT from gWAT. Then gWAT was analyzed by assessing the fat content within the peritoneal cavity using the clearly visible peritoneum as guidance. The sWAT was obtained by subtracting gWAT from total body-fat. Color-coded 3D-images of sWAT and gWAT were generated from the histograms using a volume-rendering tool (Microview software).
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