Ex vivo planar scintigraphic imaging was performed on posterior paw being removed and positioned over the collimator of a dedicated small animal Nano-SPECT-CT system (BIOSCAN™, Washington DC, USA) at 1 hour after i.v. injection of radiolabeled 111In-aptamer. The acquisition was performed during 10 minutes with two 15% windows centered on the two peaks 171 keV and 245 keV of indium 111.
Nanospect ct system
Manufactured by Bioscan
Sourced in United States, Hungary
The NanoSPECT/CT system is a preclinical multi-modality imaging device that combines single-photon emission computed tomography (SPECT) and computed tomography (CT) capabilities. The system is designed for small animal imaging, providing high-resolution functional and anatomical data.
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12 protocols using nanospect ct system
1
Ex vivo Scintigraphic Imaging of Radiolabeled Aptamer
2
Quantitative SPECT/CT Imaging of Tumor Uptake
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Forty-eight hours after injection of 177Lu-DOTATATE, helical single-photon emission computed tomography/computed tomography (SPECT/CT) scanning of the tumor region was performed with the four-headed NanoSPECT/CT system (BioScan, Washington DC, USA). Multi-pinhole rat collimators with nine pinholes (diameter 2.5 mm) per head were used: 24 projections, 90 s per projection, were applied. SPECT scans were reconstructed iteratively on a 256 × 256 matrix, using HiSPECT NG software (Scivis GmbH, Göttingen Germany) and ordered subset expectation maximization (OSEM). The total amount of radioactivity (MBq) in the tumor was quantified by drawing a sufficiently large volume of interest (VOI) around the tumor using InVivoScope software (IVS, Bioscan, Washington DC, USA). To achieve accurate quantification, the camera was calibrated by scanning a 20-ml polypropylene tube rat phantom filled with a known amount of 177Lu activity. The in vivo tumor volume was assessed by setting the lower threshold to 90% of the maximum voxel intensity of the tumor using the IRW program (Siemens, Erlangen, Germany). Before euthanasia, a whole-body SPECT/CT scan of rats was acquired 24 h after intravenous injection of 111In-DTPA-octreotide (50 MBq 111In/0.5 μg DTPA-octreotide) to detect CA20948 metastases. During scanning, the rat body temperature was maintained using a heated bed.
3
In Vivo Imaging of Anti-CD55 Antibody
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The 177Lu-anti-CD55 antibody (7.4 MBq) was injected directly into the thoracic cavity of the mice and a 30 min scan was acquired 3 and 24 hours p.i. with a NanoSPECT/CT system (Bioscan) as described previously42 (link). Images were acquired using the In Vivo Scope software (Bioscan) and PMOD v3.6.
4
Dual Receptor Imaging of Xenograft Mice
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Mice bearing dual SST2- and SST5-expressing xenografts were imaged using a nano SPECT/CT system (Bioscan, Mediso, Budapest, Hungary) 4 h after administration of [177Lu]Lu-DOTA-ST8950 (100 pmol/100 µL/6 MBq). A helical CT scan was acquired with the following parameters: current, 177 mA; voltage, 45 kVp; pitch, A helical SPECT scan was acquired using multipurpose pinhole collimators (APT1), 20% energy window width centered symmetrically over the 208 and 113 keV g-peaks of 177Lu, 24 projections, and 1200 s per projection. CT and SPECT images were reconstructed and filtered using the manufacturer’s algorithm, resulting in a pixel size of 0.3 mm for the SPECT and of 0.2 mm for the CT.
5
Quantitative SPECT/CT Imaging of Radiolabeled Tumors
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During scanning experiments, 2.0 % isoflurane/O2 gas anaesthesia was applied at 0.5 ml/min. Twenty-four hours after injection of 177Lu-TATE or 111In-octreotide, helical SPECT/CT of the tumour region was performed with a four-headed NanoSPECT/CT system (BioScan, Washington DC USA) with nine pinhole mice collimators (diameter 1.4 mm) per head. The scans were obtained using 24 projections of 120 s per projection and a quality factor of 0.7. SPECT scans were reconstructed iteratively on a 256 × 256 matrix, using HiSPECT NG software (Scivis, GmbH Göttingen Germany) and ordered subset expectation maximization (OSEM). The total amount of radioactivity (MBq) in the tumour was quantified by 3D quantification using InVivoScope software (IVS, Bioscan, Washington DC USA). To achieve accurate quantification, the camera was calibrated by scanning a 20-mL polypropylene tube phantom filled with a known amount of 177Lu or 111In radioactivity. During scanning, body temperature of the mice was maintained using a heated bed.
6
SPECT-CT Imaging of Mice
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SPECT-CT imaging was performed using a nanoSPECT/CT system (Bioscan) equipped with parallel pinhole collimators. The mice (n = 6) were anaesthetised with isofluorane, and air mixture and imaging was conducted as reported before [23 (link)].
7
Theranostic Evaluation of 177Lu-DOTA-chCE7 in Tumor-Bearing Mice
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SPECT/CT imaging studies were performed in a NanoSPECT/CT system (Bioscan, Washington, DC, USA). 177Lu-DOTA-chCE7 (6 MBq, 25 μg, 100 μl) was injected into the tail vein of tumour-bearing nude mice 24 h prior to i.p. administration of 600 μg (31.6 mg/kg) PTX or PBS. Control mice received 177Lu-DOTA-control IgG. SPECT/CT scans were performed 96 h (177Lu-DOTA-control IgG) and 120 h (177Lu-DOTA-chCE7, 177Lu-DOTA-chCE7 + PTX) after RIC administration. SPECT data were reconstructed by HighSPECT software (ver. 1.4.3049, Scivis). Reconstruction of CT data, fusion with SPECT data and analysis were performed by InVivoScope postprocessing software (ver. 1.44, Bioscan, Washington, DC, USA).
8
Radiolabeled Peptide Characterization
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AMD3100 was purchased from Toronto
Research Chemicals (Canada).
All other reagents and solvents were purchased from Acros Organics
(Belgium) and Merck (Darmstadt, Germany) and used without further
purification. DOTA-tris(tBu)ester was purchased from CheMatech (France).
Purity of the peptides was >95% as assessed by liquid chromatography
mass spectrometry (LC-MS) on a LCMS-2020 SHIMADZU (Japan) system equipped
with a Waters XBridge C-18 column (4.6 mm × 150 mm, 5 μm
particle size). The gradient used was 15–65% solvent B in 15
min (A = H2O [0.1%TFA], B = ACN [0.1% TFA]) at a flow rate of 1.0 mL/min. Radio-HPLC was performed
on an Agilent 1260 infinity instrument (Agilent) connected to a GABI
radioactivity-HPLC-flow-monitor γ-spectrometer (Elysia-raytest,
Germany). Radioligands were analyzed using Phenomenex Jupiter Proteo
C12 (90 Å, 250 mm × 4.6 mm) column using the gradient 5–50% B in 15 min (A = H2O [0.1%TFA], B = ACN [0.1% TFA]) with a flow rate of 2 mL/min. Quantitative
γ-counting was carried out on a COBRA 5003 γ-system well
counter from Packard Instruments. SPECT/CT images were acquired using
a dedicated nanoSPECT/CT system (Bioscan, Mediso, Hungary).
Research Chemicals (Canada).
All other reagents and solvents were purchased from Acros Organics
(Belgium) and Merck (Darmstadt, Germany) and used without further
purification. DOTA-tris(tBu)ester was purchased from CheMatech (France).
Purity of the peptides was >95% as assessed by liquid chromatography
mass spectrometry (LC-MS) on a LCMS-2020 SHIMADZU (Japan) system equipped
with a Waters XBridge C-18 column (4.6 mm × 150 mm, 5 μm
particle size). The gradient used was 15–65% solvent B in 15
min (A = H2O [0.1%TFA], B = ACN [0.1% TFA]) at a flow rate of 1.0 mL/min. Radio-HPLC was performed
on an Agilent 1260 infinity instrument (Agilent) connected to a GABI
radioactivity-HPLC-flow-monitor γ-spectrometer (Elysia-raytest,
Germany). Radioligands were analyzed using Phenomenex Jupiter Proteo
C12 (90 Å, 250 mm × 4.6 mm) column using the gradient 5–50% B in 15 min (A = H2O [0.1%TFA], B = ACN [0.1% TFA]) with a flow rate of 2 mL/min. Quantitative
γ-counting was carried out on a COBRA 5003 γ-system well
counter from Packard Instruments. SPECT/CT images were acquired using
a dedicated nanoSPECT/CT system (Bioscan, Mediso, Hungary).
9
Small Animal SPECT/CT Imaging
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SPECT/CT images were acquired using a dedicated nanoSPECT/CT system (Bioscan, Mediso, Budapest, Hungary). Mice were injected intravenously via the tail vein with ~ 9–15 MBq (500 pmol) of the radioligand and euthanized after 4 h. Details on image acquisition and reconstruction parameters are described in the “Supplementary information .”
10
In Vivo Imaging of 177Lu-OPS201 in Mice
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Mice were imaged using a nanoSPECT/CT system (Bioscan) 4 h after intravenous administration of 100 mL/5 MBq of 177 Lu-OPS201 (20, 200, and 2,000 pmol). A helical CT scan was acquired with the following parameters: current, 177 mA; voltage, 45 kVp; pitch, 1. A helical SPECT scan was acquired using multipurpose pinhole collimators (APT1), 20% energy window width centered symmetrically over the 208-and 113-keV g-peaks of 177 Lu, 24 projections, and 1,200 s per projection. CT and SPECT images were reconstructed and filtered using the manufacturer's algorithm, resulting in a pixel size of 0.3 mm for the SPECT and of 0.2 mm for the CT.
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