Five mice bearing U87-GFP tumours were each administered 300 μL of freshly prepared radiolabelled particles through the tail vein with a corresponding dose of 0.3 mCi per animal and 6.75 mg kg–1 porphyrin. Six hours post administration, each mouse was euthanized using cardiac puncture. A complete tissue biodistribution was performed inclusive of all major organs, tumour, peri-tumoural area and normal brain tissue from the contralateral hemisphere. Each collected tissue and blood was weighed and measured for 64Cu gamma-activity using an automatic gamma counter (Perkin Elmer, Waltham, MA, USA) with a linear detection limit for 64Cu of 1.88 nCi (60 second integration time). Activity measurements were decay-corrected to the time of injection and expressed as a percentage of the injected dose per gram of tissue.
Automatic gamma counter
Manufactured by PerkinElmer
Sourced in United States
The Automatic Gamma Counter is a laboratory instrument designed to measure the radioactivity of samples. It accurately quantifies the amount of gamma radiation emitted by radioactive isotopes present in the samples. The device automatically processes multiple samples, providing efficient and precise measurements of radioactivity levels.
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13 protocols using automatic gamma counter
1
Biodistribution of Radiolabelled Particles
2
PET/CT Imaging of Candida Infection in Mice
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Three mice from each group were anaesthetized by intraperitoneal injection of 1% pentobarbital and tails were intravenously injected with [18F]FDG (Guangzhou HTA Isotope Medical Co. Ltd) (10–11 MBq) at 24, 48 and 72 h after infection with C. albicans (5 × 105 CFU). Forty minutes after the injection, the mice were scanned with a small-animal PET/CT scanner (Inveon) for 20 min. All PET images were reconstructed with the maximum a posteriori (MAP) reconstruction algorithm in Inveon Acquisition Workspace (Siemens Medical Solutions Inc.) and analysed with Inveon Research Workspace 4.2 (Siemens Medical Solutions Inc.) after the coregistration of the PET/CT images. Regions of interest (ROIs) were drawn in the kidneys. The radioactivity density from image analysis is presented as the percent injected dose per gram (% ID/g). After the final imaging time point, the mice were sacrificed. The blood, brain, heart, lungs, liver, spleen, kidneys, stomach, colon and muscle were harvested for ex vivo biodistribution analysis. The radioactivity in each organ was measured with an automatic gamma counter (Perkin-Elmer) after which the organ weights were determined with a balance. The biodistribution is presented as a percent of the injected dose per gram (% ID/g)45 (link),46 (link).
3
Measuring GFR Using 51Cr-EDTA
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GFR was measured as urinary 51Cr-EDTA clearance. A bolus of 51Cr-EDTA was followed by continuous intravenous infusion. Blood and urine samples were collected at regular intervals as described above and counted in a gamma counter (Perkin Elmer 2480, Automatic Gamma Counter, Wizzard). Correction for background and decay was performed.
4
LPS-Induced Neuroinflammation in Mice
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ICR male mice aged between 5 and 6 weeks were used in this study. The mice were provided drinking water and a normal diet ad libitum and were maintained under a 12 h light-dark cycle at 24 ± 1 °C with 50% humidity. The LPS-treated mouse model was prepared by injecting LPS (10 mg/kg in saline) i.p. into mice, and all experiments were carried out on the third day after LPS injection. Tissue radioactivity was counted using an automatic gamma counter (PerkinElmer, Waltham, MA, USA). MicroPET/CT images were acquired using an Inveon microPET/CT scanner (Siemens Medical Solutions, Knoxville, TN, USA).
All animal experiments were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee of Samsung Medical Center (20210104001).
All animal experiments were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee of Samsung Medical Center (20210104001).
5
Peptide Partitioning Protocol
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Labeled peptide was diluted with 600 μL of phosphate-buffered saline (PBS), and 50 µL of the solution was mixed in the test tube with 450 μL of PBS and 500 μL of octanol. The sample was placed on the shaker (1000 RPM) for 20 min and then centrifuged at 15,000 RPM for 1 min. Subsequently, 50 µL from both octanol and water layer were collected and measured on an automatic gamma counter (PerkinElmer, Waltham, Massachusetts, USA). Partition coefficient was calculated as the decadic logarithm of the ratio of octanol/water activity.
6
Cellular Uptake of Radiolabeled Nectin-4 Antibody
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Seed the cells were in 24-well plates with a cell concentration of 1.0 × 105 cells/well. After complete attachment, replace 1 mL serum-free medium containing 99mTc-Hynic-mAbNectin-4 (37 kBq) to each well and incubation was performed at 37 °C for multiple time points (1, 2, 3, 4,6, and 8 h). For each time point, rinse the radiotracer-treated cells with 800 μL pre-cooling PBS twice. Collect the rinsed PBS as supernatants. Then 800 µL NaOH (1 N) was added for cell lysis and collected as lysates. Radioactivity of the supernatants and lysates was detected using automatic gamma-counter (PerkinElmer, USA). After attenuation correction, the cell uptake rate is determined as: Alysate/(A supernatant + Alysate) × 100%. For blocking studies, experimental cells were incubated with excess unradiolabeled mAbNectin-4 for 1 h before experiments.
7
In Vivo PET Imaging of Micellar Biodistribution
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All animal studies
were conducted under a protocol approved by the University of California,
Davis, Animal Care and Use Committee. 64CuCl2 was purchased from Washington University (St Louis, MO). The details
of the synthesis of radiolabeled micelles and PET imaging scans are
described in previous work.18 (link) In brief,
the pharmacokinetics and in vivo biodistribution
of C16- and C18-micelles were studied by positron emission tomography
(PET) and an automatic gamma counter (PerkinElmer, CT), respectively,
after intravenous administration of the micellar solution to female
FVB mice (n = 6 for C18-micelles, (371 ± 107
μCi and 0.68 ± 0.15 mg per mouse) and n = 4 for C16-micelles, (407 ± 9 μCi and 0.69 ± 0.008
mg/mouse) bearing new deletion mutant (NDL)35 (link),36 (link) tumors implanted bilaterally within the mammary fat pads. All PET
images and the organ distribution of 64Cu-labeled micelles
presented here have been decay corrected.
were conducted under a protocol approved by the University of California,
Davis, Animal Care and Use Committee. 64CuCl2 was purchased from Washington University (St Louis, MO). The details
of the synthesis of radiolabeled micelles and PET imaging scans are
described in previous work.18 (link) In brief,
the pharmacokinetics and in vivo biodistribution
of C16- and C18-micelles were studied by positron emission tomography
(PET) and an automatic gamma counter (PerkinElmer, CT), respectively,
after intravenous administration of the micellar solution to female
FVB mice (n = 6 for C18-micelles, (371 ± 107
μCi and 0.68 ± 0.15 mg per mouse) and n = 4 for C16-micelles, (407 ± 9 μCi and 0.69 ± 0.008
mg/mouse) bearing new deletion mutant (NDL)35 (link),36 (link) tumors implanted bilaterally within the mammary fat pads. All PET
images and the organ distribution of 64Cu-labeled micelles
presented here have been decay corrected.
8
Comprehensive Biodistribution Analysis
9
Cellular Uptake of Radiolabeled Nectin-4 Antibody
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Seed the cells were in 24-well plates with a cell concentration of 1.0 × 105 cells/well. After complete attachment, replace 1 mL serum-free medium containing 99mTc-Hynic-mAbNectin-4 (37 kBq) to each well and incubation was performed at 37 °C for multiple time points (1, 2, 3, 4,6, and 8 h). For each time point, rinse the radiotracer-treated cells with 800 μL pre-cooling PBS twice. Collect the rinsed PBS as supernatants. Then 800 µL NaOH (1 N) was added for cell lysis and collected as lysates. Radioactivity of the supernatants and lysates was detected using automatic gamma-counter (PerkinElmer, USA). After attenuation correction, the cell uptake rate is determined as: Alysate/(A supernatant + Alysate) × 100%. For blocking studies, experimental cells were incubated with excess unradiolabeled mAbNectin-4 for 1 h before experiments.
10
Biodistribution and PET Imaging of Radiolabeled IMB1636
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Mice were euthanized after the terminal time point of PET and SPECT imaging. Tumors and major organs including heart, blood, liver, spleen, lung, kidney, stomach, intestine, pancreas, bladder, muscle, bone, brain, and skin were all collected and weighed for biodistribution study. An automatic gamma counter (PerkinElmer) was used to measure the radioactivity counts. Biodistribution data was presented as %ID/g. 18 F-FDG PET Imaging 18 F-FDG PET imaging was performed at 14 days to evaluate the RIT therapeutic effect in high-177 Lu-IMB1636 and 177 Lu-only groups. The mice were kept fasting for at least 6 h before PET imaging. Each mouse was intravenously injected with ~7.4 MBq of 18 F-FDG and was anesthetized with iso urane for about 40 ~ 60 minutes before PET imaging. Quantitative analysis was performed by drawing ROI of tumor.
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