8–10 week old C57BL/6 mice were injected i.v. with LLC exosomes (10μg, in 100μl PBS) every 3 days for 2 weeks. Mice were fasted for 12 hours prior to intraperitoneal injection with 18FDG. One hour after injection, the mice were euthanized, lungs were harvested and measured on a Biodex Atomlab™ 500 for radioactivity. Lung tissue was enzymatically digested (collagenase (5g/L), Hyaluronidase (0.4g/L), DNAse I (0.15g/L)) for 20 minutes with rotation at 37°C. Following digestion, RBC’s were lysed using ACK. Cells were stained with primary Biotin anti-mouse CD19 (Biolegend,115504), secondary Streptavidin MicroBeads (Miltenyl Biotec,130-048-101), CD8a (Ly-2) MicroBeads (Miltenyl Biotec,130-049-401), CD4 (L3T4) (Miltenyl Biotec,130-049-201) for 15 minutes. Cells were then magnetically separated on an autoMACS Pro separator (Miltenyl Biotec) using the positive selection protocol. The negative fraction was then collected and read on a Biodex Atomlab™ 500 for radioactivity. Cells were then resuspended in Trizol and saved in the −80°C for RT-PCR.
Atomlab 500
Manufactured by Mirion Technologies
Sourced in United States
The Atomlab 500 is a laboratory instrument designed for the measurement of radioactivity. It is a versatile and accurate device used for the analysis of various samples, including environmental, medical, and industrial materials. The Atomlab 500 provides precise measurements of radioactive isotopes and their concentrations, enabling users to effectively monitor and assess radiation levels.
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Lab products found in correlation
Biotin anti mouse cd19, by BioLegend (1 mentions)
Cd4 l3t4, by Miltenyi Biotec (1 mentions)
Automacs pro separator, by Miltenyi Biotec (1 mentions)
Streptavidin microbeads, by Miltenyi Biotec (1 mentions)
Cd8a ly 2 microbeads, by Miltenyi Biotec (1 mentions)
Topotecan hydrochloride hydrate, by Merck Group (1 mentions)
Solid phase extraction cartridges, by Waters Corporation (1 mentions)
N n dimethylformamide dmf anhydrous 99.8, by Merck Group (1 mentions)
Packard cobra 2 auto gamma counter, by PerkinElmer (1 mentions)
Gemini c18, by Phenomenex (1 mentions)
8 protocols using atomlab 500
1
Quantification of Tumor-Derived Exosome Uptake
2
Re-entrant Chamber Quality Assurance
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The nuclear medicine department’s QA program for the re-entrant chamber was evaluated. The department performs daily constancy checks to ensure the accuracy of activity measurements of the re-entrant chamber. Linearity check was performed quarterly (more frequently if indicated by the constancy check) by measuring the dose calibrator’s response to a series of sources with different activities and plotting them (18 , 19 (link)). A sensitivity analysis measured the chamber’s response to different source geometries. An authorized physicist independently reviewed these results regularly. AtomLab™ 500 (BIODEX, Mirion Technologies, NJ, USA) re-entrant chambers were used for Lutathera and Pluvicto measurements calibrated with a National Institute of Standards and Technology (NIST) traceable dose provided by the drug manufacturer. The dose measurement accuracy was verified on multiple days against decayed activity.
3
Validation of SPECT Imaging Protocols
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All static and WB planar measurements were acquired with a single detector only, as the gamma camera acceptance tests showed little difference between the system's two detectors. The amount of radioactivity used was sufficiently low that the camera did not display any dead-time effects; thus, no dead-time correction was necessary. Before measurements, the detector was peaked relative to the 159.0 keV and 364.5 keV photopeak's for all I-123 and I-131 measurements, respectively. SPECT measurements were, however, conducted using both detectors to increase sensitivity for a given acquisition time.
I-123 and I-131 activities were measured using a Biodex Atomlab 500 dose calibrator (Biodex Medical Systems, New York, NY, USA). The accuracy of the dose calibrator for both I-123 and I-131 was traceable to a secondary standard through the National Institute of South Africa (NMISA) in Cape Town, South Africa.
CT images were acquired for each static planar, WB planar and SPECT validation test setup, for use in both the SIMIND simulation and in the reconstruction processes, as explained below. The CT images were acquired in a standard imaging protocol (image matrix: 512 × 512, pixel size: 1.2 × 1.2 mm2, slice thickness: 5.0 mm, reconstruction kernel: standard smoothing body kernel (B08s)).
I-123 and I-131 activities were measured using a Biodex Atomlab 500 dose calibrator (Biodex Medical Systems, New York, NY, USA). The accuracy of the dose calibrator for both I-123 and I-131 was traceable to a secondary standard through the National Institute of South Africa (NMISA) in Cape Town, South Africa.
CT images were acquired for each static planar, WB planar and SPECT validation test setup, for use in both the SIMIND simulation and in the reconstruction processes, as explained below. The CT images were acquired in a standard imaging protocol (image matrix: 512 × 512, pixel size: 1.2 × 1.2 mm2, slice thickness: 5.0 mm, reconstruction kernel: standard smoothing body kernel (B08s)).
4
Topotecan Quantification Protocol
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Cassettes, reagent kits (Prod No. PE-FSPG-047-R), the precursor (Prod No. 3193.0075), elution solution (Prod No. PE-FSPG-047-R-V1), and reference standard (PE-FSPG-047-H) were purchased from ABX (Radeberg, Germany). N-Desmethyl topotecan was purchased from Synfine Research (Toronto, Canada). Topotecan hydrochloride hydrate (>98%) standard and N,N-dimethylformamide (DMF, anhydrous 99.8%) were purchased from Sigma-Aldrich (St. Louis, MO, USA) and used without further purification. Sodium hydroxide National Institute of Standards and Technology (NIST) traceable solution (0.5 N) was purchased from Aqua Solutions, Inc. (Deer Park, TX, USA) and used without further purification. Solid-phase extraction cartridges were purchased from Waters (Milford, MA, USA). Ultra-high purity N.O.S. gas (99% nitrogen/1% oxygen) was purchased from Airgas (White Plains, NY, USA). All other reagents not listed above were of the highest grade available from Sigma-Aldrich (St. Louis, MO, USA) and Fisher Scientific (Pittsburgh, PA, USA). End of synthesis radioactivity was determined using a Biodex AtomLab 500 dose calibrator (Shirley, NY, USA).
5
Radiopharmaceutical Cell Labeling Stability
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Stability of labelled cells was assessed incubating granulocytes, lymphocytes, and platelets in PBS at 37°C. After 1 h and 3 h, an aliquot from each vial was centrifuged to collect pellet and supernatant that were counted for radioactivity with a single-well gamma counter (Atomlab 500, Biodex) in order to evaluate the radiopharmaceutical elution from labelled cells over time.
The trypan blue exclusion test was also performed in order to verify the viability of each cell population at different time points after labelling.
The trypan blue exclusion test was also performed in order to verify the viability of each cell population at different time points after labelling.
6
Biodistribution of Gallium-67 Tracer
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A full biodistribution was routinely conducted (blood, heart muscle, liver, kidneys, lungs, small intestine, brain, bladder, muscle, spleen, stomach, bone, tumor, and pancreas) after the last scan on day 8 (192 h). Organs were cleaned from blood, weighed and the activity determined using a γ-counter (Packard Cobra II auto-gamma counter, Perkin Elmer, Waltham, MA, USA). The calibration factor for 37 kBq of 67Ga was 1 524 228 cpm (instrument specific). High activity organs (blood, liver) were measured in an Atomlab 500 dose calibrator (Biodex, Shirley, NY, US) if the activity exceeded 37 kBq. Total organ weights were used for the calculations of injected dose per organ (% ID per g organ) except for blood, liver, muscle, bone and pancreas, where average literature values were used.43–45 (link)
7
Synthesis and Characterization of 9-(4-18F-Fluoro-3-Hydroxymethyl)Butyl Guanine
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Fluorine-18 was obtained from VOXEL S.A. (Cracow, Poland) with radiochemical purity > 99.9%. The input activity per synthesizer was 10 GBq. Product quality control comprised testing radiochemical and chemical purity. High-performance liquid chromatography (HPLC) (Shimadzu AD 20 HPLC with UV–Vis detector) with a radiometric detector (GabiStar, Raytest, Germany) was used to determine these parameters. During HPLC analysis, a C18-RP (Phenomenex Gemini C18 150 mm × 4.6 mm × 5 µm) column was used. The Atomlab 500 (Biodex Medical Systems, Shirley, NY, USA) dose calibrator was used for all activity measurements. 9-(4-18F-fluoro-3-[hydroxymethyl]butyl)guanine was synthesized at the University of Warsaw, Biological and Chemical Research Centre. We obtained the product with a radiochemical purity > 98.5% (Supplementary Fig. S9 ).
8
Lutetium-177 SPECT-CT Imaging Protocol
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The radionuclide Lutetium-177 (177-Lu), which is widely used in systemic treatments, was used for the scans. Biodex Atomlab 500 (Biodex Medical Systems New York, NY, USA) dose calibrator certified by the accredited company was used for radionuclide dose measurements. GE Discovery NM 670 SPECT-CT (GE Medical Systems, Waukesha, WI, USA) device and medium-energy general-purpose collimator (MEGP) were used in the scans. In line with the EANM/MIRD Guidelines(10 (link), 30 (link)), during the SPECT acquisitions 60 projections, 20 second/projection, 128 × 128 matrix size, body contour on parameters were used.
The images were then reconstructed using Ordered Subset Expectation Maximisation (OSEM) with five iterations, 10 subsets and no post-filter as done in the study of Wevrett et al.(25 ). During the CT acquisitions, 120-kV and 200-mAs parameters were used in line with the user manual recommendation of CT. Then, all phantom images were reconstructed without and with CT-based scatter and attenuation correction as described below.
The images were then reconstructed using Ordered Subset Expectation Maximisation (OSEM) with five iterations, 10 subsets and no post-filter as done in the study of Wevrett et al.(25 ). During the CT acquisitions, 120-kV and 200-mAs parameters were used in line with the user manual recommendation of CT. Then, all phantom images were reconstructed without and with CT-based scatter and attenuation correction as described below.
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