Male DBA/1JRj mice (starting weight 24.3 ± 0.8 g (n = 10/group)) with CIA were randomly divided between the treatment groups upon development of overt arthritis (macroscopic score of inflammation > 0.5). Upon inclusion, the mice were injected intravenously with 5 µL liposomes (in 200 µL PBS) with or without 1 µg IRDye700DX and labeled with 0.6 MBq 111In for biodistribution analysis. A subset of mice (n = 2 per group) were injected with 18 MBq 111In for SPECT/CT imaging. After 24 h, the mice were sacrificed, and the relevant tissues were dissected and weighed. Tissue uptake of 111In was determined using a γ counter (WIZARD, 2480 Automatic Gamma Counter, Perkin Elmer, Waltham, MA, USA). Results are depicted as percentage of the injected amount per gram tissue (%IA/g). Mice which received a SPECT/CT dose of 111In were scanned for 60 min (4 × 15 min frames) using a 1-mm-diameter pinhole ultra-high sensitivity mouse collimator (U-SPECT/CT-II, MILabs, Houten, The Netherlands). SPECT scans were followed by CT scans (65 kV, 615 µA). The SPECT scans were reconstructed using software from MILabs using a 0.2 mm voxel size, 1 iteration and 16 subsets.
U spect 2 ct
Manufactured by MILabs
Sourced in Netherlands
The U-SPECT-II/CT is a small-animal imaging system designed for preclinical research. It combines high-resolution SPECT (Single-Photon Emission Computed Tomography) and CT (Computed Tomography) imaging modalities to provide comprehensive anatomical and functional data. The system is capable of acquiring 3D tomographic images of small animals such as mice and rats.
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12 protocols using u spect 2 ct
1
Biodistribution and SPECT/CT Imaging of Liposomal Delivery in CIA Mice
2
SPECT/CT Imaging in Rodent Studies
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SPECT/CT scans were acquired for either 1
h at 24 or 48 h post injection or for 1.5 h at 96 h post injection
using a U-SPECT/CT-II (MILabs, Utrecht, The Netherlands). Images were
acquired using a 1 mm diameter pinhole ultrahigh sensitivity mouse
collimator or a 1 mm diameter pinhole rat collimator (n = 1). SPECT scans were followed by CT scans (65 kV, 615 μA).
All SPECT scans were reconstructed with 3 iterations and 16 subsets
and a voxel size of 0.4 mm (MILabs reconstruction software). SPECT
images were made with VivoQuant software. The tissues from the mice
that underwent the SPECT/CT scans were also used for the ex vivo biodistribution
as described above after scanning.
h at 24 or 48 h post injection or for 1.5 h at 96 h post injection
using a U-SPECT/CT-II (MILabs, Utrecht, The Netherlands). Images were
acquired using a 1 mm diameter pinhole ultrahigh sensitivity mouse
collimator or a 1 mm diameter pinhole rat collimator (n = 1). SPECT scans were followed by CT scans (65 kV, 615 μA).
All SPECT scans were reconstructed with 3 iterations and 16 subsets
and a voxel size of 0.4 mm (MILabs reconstruction software). SPECT
images were made with VivoQuant software. The tissues from the mice
that underwent the SPECT/CT scans were also used for the ex vivo biodistribution
as described above after scanning.
3
In Vivo SPECT/CT Imaging of Antibodies
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For each construct, mice (n = 5/group) received 0.4 pmol 111In-labelled tracer (8 MBq, 200 µL). For Fab, scans were acquired at 4 h and 24 h post-injection (p.i.), for Fab-PEG at 4 h, 24 h and 48 h, and the full length antibodies were imaged at 24 and 72 h p.i.. Images were acquired for 45 min under general anesthesia (isoflurane in 100% oxygen, 5% for induction, 2% maintenance) with the U-SPECT-II/CT (MILabs, Utrecht, The Netherlands) using a 1.0 mm diameter pinhole mouse high sensitivity collimator, followed by CT scan (615 µA, 65 kV) for anatomical reference. Scans were reconstructed with MILabs reconstruction software using a 16-subset expectation maximization algorithm, with a isotropic voxel size of 0.2 mm and 1 iteration. SPECT/CT scans were analyzed and maximum intensity projections (MIP) were created using Inveon Research Workplace software (Siemens).
4
Iodinated Bone Imaging in Fracture Mice
5
In Vivo Tumor Imaging with 111In-F4/80
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In microSPECT imaging studies, mice with orthotopic MDA-MB-231 tumour xenografts were injected i.v. with 100 μg purified 111In-F4/80 (14 MBq). Mice were euthanized at 24 h p.i. and scanned using the U-SPECT-II/CT (4 frames of 15 min; spatial resolution 160 μm, 65 kV, 615 μA) (MILabs) [30 (link)]. A CT scan was also taken for anatomic reference. Mice were euthanized prior to imaging, rather than imaged under anaesthesia, to allow a proper comparison of the SPECT/CT imaging data with the biodistribution data. SPECT scans, four frames combined, were reconstructed using an ordered subset expectation maximization algorithm, with a voxel size of 0.4 mm (MILabs).
6
In Vivo Renal Function Monitoring
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To monitor the renal function in vivo, mice in the tolerability study underwent SPECT imaging using 99mTc-labeled dimercaptosuccinic acid (DMSA) at baseline, 10 weeks p.i., and 16 weeks p.i. Preparation of [99mTc] Tc-DMSA (99mTc-DMSA) was performed as per the manufacturer’s protocol (Curium Netherlands B.V., Petten, the Netherlands). In short, 740–1100 MBq of pertechnetate was added to 1.2 mg of DMSA and incubated for 15 min at 25 °C. Mice received an intravenous tail injection of 20 MBq 99mTc-DMSA in 200 µL saline 2 h prior to SPECT imaging. The SPECT scans were acquired with the U-SPECT-II/CT (MILabs, Utrecht, the Netherlands) using a 1.0 mm diameter pinhole mouse high sensitivity collimator and acquisition time of 15 min. Image reconstruction was performed with MILabs reconstruction software using a 16-subset expectation maximization algorithm with a voxel size of 0.2 mm and 3 iterations. Quantification of renal uptake of 99mTc-DMSA was performed by drawing volumes of interest (VOIs) around the kidneys. The radioactivity was corrected for decay and volume of the VOI, which resulted in a percent injected activity per gram kidney tissue, assuming a tissue density of 1.0 g/cm3.
7
Skeletal Tissue Imaging via Micro-SPECT/CT
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Mice were scanned using a U-SPECT-II/CT (MILabs) under general anesthesia (isoflurane/O2) for 15 min using the 1.0-mm diameter pinhole mouse high sensitivity collimator tube, followed by a CT scan (spatial resolution 160 mm, 65 kV, 615 mA) for anatomical reference. Scans were reconstructed with MILabs reconstruction software using an ordered-subset expectation-maximization algorithm, with a voxel size of 0.4 mm. SPECT/CT scans were analyzed and maximum intensity projections were created using the Inveon Research Workplace software (IRW, version 4.1). A three-dimensional (3D) volume of interest (VOI) was drawn using CT threshold (CT value: soft tissue is 11–39% and skeletal tissue is 41–100%) to differentiate soft tissue from skeletal tissue, and uptake was quantified as the percentage injected dose per gram (%ID/g) using standard curve, assuming a tissue density of 1 g/cm3. The hot spot in the skeletal tissue ROI was chosen with the location of the edge of the ROI contour representing 75% of maximum intensity. All mice were euthanized with CO2 after micro-SPECT/CT imaging.
8
SPECT/CT Imaging of Tumor-Bearing Mice
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For SPECT/CT imaging, HeLa xenograft-bearing mice were anesthetized with 0.8%–1.8% isoflurane in air; tumor-bearing mice (n=3) received intratumoral injections of 15 MBq of either 111In-MNT-PEG-FA or control 111In in Hanks solution in a volume equal to half of the tumor volume. Whole-body imaging was performed on a U-SPECT-II/CT (MILabs, Utrecht, the Netherlands) scanner immediately after injection and continued for 5×10-min frames using a 1.0-mm-diameter pinhole collimator, with subsequent immediate whole-animal CT acquisition. Additional SPECT/CT imaging was performed during the subsequent days (3–5 frames ×10 min). The images were reconstructed using U-SPECT-Rec2.34b software obtained from the manufacturer, followed by co-registration of SPECT images to the corresponding CT images. Quantitative analysis of images after three dimensional (3D)-reconstruction was performed using PMOD 3.4 software (PMOD Technologies Ltd., Zurich, Switzerland).
The radioactivity concentration in tumor and other organs or tissues was determined by selecting a few spheres within the organ or tissue of interest, followed by division of the summarized signal within this sphere by its volume.
The radioactivity concentration in tumor and other organs or tissues was determined by selecting a few spheres within the organ or tissue of interest, followed by division of the summarized signal within this sphere by its volume.
9
Imaging Tibial Lesions with 99mTc-medronate
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99mTc-medronate was prepared as per the protocol provided by MDP multidose kit (GE Healthcare, The Netherlands). All mice received an intravenous injection with a dose of approximately 37 MBq 99mTc through the tail vein. Immediately after 1 hpostinjection, images were acquired using a U-SPECT-II/CT (MILabs), as reported previously [15 (link),17 (link),18 ]. The following criteria were used to confirm the tibial lesion formation: a) visual confirmation of 99mTc-uptake below the growth plate in right tibia compared to contralateral control tibia; b) at least 10% 99mTc-uptake increase in tibial lesion region of interest (ROI) below growth plate compared to contralateral control tibia.
10
In Vivo SPECT/CT Imaging of Tumor-Targeting Radionuclide
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For SPECT/CT imaging, the animals were anesthetized with 0.8%–1.8% isoflurane in air. The mice (n=4; mean tumor volume: 88±19 mm3) received intratumoral bolus injections of 7.3±1.1 MBq of 111In-NOTA–MNT-MSH in 45 μL. Whole body imaging was performed on an U-SPECT-II/CT (MILabs, Utrecht, the Netherlands) scanner, beginning immediately after injection and continuing for 2.5 h (13×10-min frames) using a 1 mm diameter pinhole collimator, with subsequent immediate whole-animal CT acquisition. Additional SPECT/CT imaging was performed up to 8 days (5 frames ×10 min). The images were reconstructed using U-SPECT-Rec2.34b software obtained from the manufacturer, followed by co-registration of SPECT images to corresponding CT images. Quantitative analysis of images after three-dimensional reconstruction was performed using PMOD 3.4 software (PMOD Technologies Ltd., Zürich, Switzerland).
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