Freshly dissected, whole rat OE was removed and submerged in OCT mounting medium (25608-930; VWR) followed by snap freezing on dry ice. Frozen, embedded tissue was sliced in 10-μm coronal sections onto slides (4 sections per slide) using a cryostat (HM550; Thermo Fisher Scientific). Slides were immersed in 4% PFA in phosphate buffer (BM-698, Boston Bioproducts) containing 2% EtOH for 30 minutes followed by 10 mM Tris, pH 7.5, at 4°C for 10 minutes. Sections were then submerged for 20 minutes in 10 mM Tris buffer containing nonradiolabeled GV1-57 (10, 5, 0.5, 0.1, 0 μM) and 5% DMSO. To these solutions was added [11C]GV1-57 (~200 nCi), before incubation at RT for 12 minutes. Slides were then washed in 10 mM Tris buffer for 10 minutes at RT and dried under vacuum for 30 minutes at 25°C. All slides were exposed to multisensitive phosphor screens (PerkinElmer) for 1 hour and imaged with a Cyclone Plus Storage Phosphor system (PerkinElmer). Images were colored using the Rainbow lookup table in ImageJ (NIH) with equivalent thresholds for brightness. Intensity values for each OE slice were measured using ImageJ, background subtracted, and averaged across replicate sections. All values were normalized to those of the DMSO-treated OE sections and displayed using GraphPad Prism software.
Multisensitive phosphor screen
Manufactured by PerkinElmer
Sourced in United States
Multisensitive phosphor screens are a type of laboratory equipment used to detect and measure various types of radiation. They are designed to be sensitive to a wide range of radiation, including X-rays, gamma rays, and charged particles. The screens are coated with a phosphor material that emits light when exposed to radiation, and the intensity of the light emission is proportional to the amount of radiation detected. This allows the screens to be used for quantitative measurements of radiation levels.
Automatically generated - may contain errors
Lab products found in correlation
Cyclone plus storage phosphor system, by PerkinElmer (3 mentions)
Optiquant, by PerkinElmer (2 mentions)
Oct mounting medium, by Avantor (1 mentions)
Hm550, by Thermo Fisher Scientific (1 mentions)
Bm 698, by Boston BioProducts (1 mentions)
Cyclone plus phosphor imager, by PerkinElmer (1 mentions)
Silica gel 60f 254 nm, by Merck Group (1 mentions)
Cr 35 bio, by Raytest (1 mentions)
Typhoon trio, by GE Healthcare (1 mentions)
Immobilon, by Merck Group (1 mentions)
Gen5 microplate reader and imager software, by Agilent Technologies (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Protease inhibitor, by Merck Group (1 mentions)
Typhoon 8600 phosphorimager, by GE Healthcare (1 mentions)
Optiquant software, by PerkinElmer (1 mentions)
10 protocols using multisensitive phosphor screen
1
Autoradiographic Quantification of Olfactory Receptor Binding
2
Tc-99m-DTPA-can225IgG Binding Assay
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10 μm thick, untreated frozen canine mammary carcinoma sections were thawn for 5 minutes at RT and then blocked in TBS+2% bovine serum albumin (BSA) for 30 min. The slides were then incubated with ∼30 kBq/section 99mTc-DTPA-can225IgG for 1h at RT. Subsequently, slides were incubated 2x for 5 min in ice-cold TBS+2% BSA and then dipped 10x into ice-cold H2O followed by drying under a cold air stream. Dry slides were placed on multisensitive phosphor screens (cat# 7001724, Perkin Elmer) for 24h and then recorded in a Cyclone Plus phosphor imager and analyzed using the OptiQuant® software (Perkin Elmer).
3
Quantifying Metoclopramide Metabolism in Animals
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Different animal groups (Table I ) were i.v. injected under isoflurane/air anesthesia with [11C]metoclopramide (28 ± 12 MBq, corresponding to 0.4 ± 0.4 μg of unlabeled metoclopramide). At 15 min after radiotracer injection, blood was collected from the retro-bulbar plexus and animals were killed by cervical dislocation while under deep anesthesia. Blood was centrifuged to obtain plasma, the liver and the kidneys were removed and urine was collected. Proteins were precipitated by the addition of acetonitrile (1 μL/μL plasma; 1000 μL for the liver, 200 μL for the kidneys and 0.5 μL/μL urine). All solutions were vortexed and centrifuged. Each supernatant (plasma, liver, kidneys and urine, 5 μL each) and diluted [11C]metoclopramide solution as a reference were spotted on thin-layer chromatography (TLC) plates (silica gel 60F 254 nm, 10 × 20 cm; Merck, Darmstadt, Germany) and the plates were developed in ethyl acetate/ethanol/ammonium hydroxide (25%, w/v) (80/20/5, v/v/v). Detection was performed by placing the TLC plates on multisensitive phosphor screens (PerkinElmer Life Sciences, Waltham, MA). The screens were scanned at 300 dpi resolution using a PerkinElmer Cyclone® Plus Phosphor Imager (Perkin-Elmer Life Sciences). The retardation factor (Rf) for [11C]metoclopramide was 0.6, while the radiolabeled metabolites remained on the start (Rf = 0).
4
Evaluating Renal Function Biomarkers
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For g-counting, plasma was collected via the tail artery during the PET acquisition (10 min after tracer delivery). After the PET acquisition (35 min after tracer injection), the rats were euthanized, urine was collected by opening the abdominal cavity, and autoradiography studies were conducted. Plasma (4 ARF rats, 4 controls) and urine (3 ARF rats, 3 controls) were counted for radioactivity in an automated g-counter (Wizard; PerkinElmer). The radioactivity concentrations, expressed as percentage injected dose per gram, were calculated (3). The kidneys were frozen and cut into 20-mm short-axis slices using a cryostat (Leica). Dual-radiotracer autoradiography was performed to assess both 18 F-FDS and 99m Tc-DTPA uptake. First, the imaging plate (MultiSensitive Phosphor Screens; PerkinElmer) was exposed for 3 h to visualize the distribution of 18 F-FDS with a digital autoradiography system (CR 35 Bio; Raytest). After 18 h to allow for the decay of 18 F, a second exposure for 12 h was started to evaluate 99m Tc-DTPA uptake. After autoradiography, tissue slices were stained with hematoxylin and eosin.
5
Quantitative In Vitro Autoradiography of [123I]FP-CIT
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For phosphor imaging autoradiography of [123I] FP-CIT animals were sacrificed at the end of imaging studies by cervical dislocation. The brain was surgically removed and immediately frozen in liquid nitrogen. Thereafter, consecutive 20 µm coronal brain sections were cut on a cryostat set to -20°C and sections were immediately apposed to a MultiSensitive Phosphor Screens (PerkinElmer, Waltham, MA, USA) for 15 minutes at room temperature. The screens were then removed from the cassettes under dim lighting and scanned at 600 dpi resolution using a Cyclone storage phosphor system. In vitro autoradiography was performed on 20-µm coronal brain sections. The amount of binding was calculated in digital light units [DLU/mm2] using the Optiquant v4.00 software (18, 19) . Semiquantitative analysis was performed within two ROI, one inside each striatum and one outside (the cerebral cortex).
Striatum-cortex/cortex ratios were calculated to evaluate specific binding. Right and left striatum were averaged, like in vivo.
Striatum-cortex/cortex ratios were calculated to evaluate specific binding. Right and left striatum were averaged, like in vivo.
6
Dual-Tracer Autoradiographic Imaging of Rabbit Heart
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In healthy New Zealand White rabbits weighing 3.8–4.2 kg (n = 4), ex-vivo high resolution autoradiographic imaging was conducted to determine accurate ventricular distribution of 18F-LMI1195. As a reference, the myocardial perfusion marker 201Tl was co-injected for dual-tracer analysis. 18F-LMI1195 (25 MBq) was injected via ear vein followed by 201Tl (0.5 MBq). Rabbits were euthanized 20 min after 18F-LMI1195 administration, and the heart was removed. The heart was sliced into 20 μm thickness slices for autoradiographic analysis. The short axis sections were exposed to a phosphor imaging plate (MultiSensitive phosphor screen; PerkinElmer, Shelton, CT) to determine 18F-LMI1195 distribution (exposure for 3 h) with a digital autoradiography system (CR 35 Bio, Raytest or Cyclone; Packard; Straubenhardt, Germany). After complete 18F decay of 3 days, sections were again re-exposed to determine 201Tl distribution (second exposure for 7 d). Figure 1 displays the described experimental setup.![]()
Schematic diagram illustrating the protocol for the dual-radiotracer autoradiography analysis using 18F-LMI1195 and 201Tl.
7
Radiolabeled MMP Probe Biodistribution in ApoE-/- Mice
8
Protein Expression and Imaging of αVβ3 Integrin
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mMSCs and human umbilical vein endothelial cells (HUVEC, known to express αVβ3) were washed three times in sterile phosphate-buffered saline (PBS). Cell pellets were collected by centrifugation and incubated in a solution of RIPA buffer (Sigma-Aldrich, USA) and Protease Inhibitor (Sigma-Aldrich, USA). The cocktail was sonicated 3 times for a length of 20 seconds with a 10 second incubation on ice. The lysed cells were centrifuged for 15 min and the supernatant (protein) was collected. The concentration of protein was determined using Gen5 Microplate Reader and Imager Software (BioTek Instruments, USA). A total of 20-10 ng of proteins were resolved on 10% SDS-polyacrylamide gel electrophoresis and transferred onto PVDF membranes (Immobilon, Millipore). Membranes were blocked in Tris-buffered saline (TBS) containing 10% non-fat dry milk and 0.1% Tween 20, prior to incubation with 80 µCi of 64Cu-NOTA-PEG4-cRGD2 targeted to αVβ3 (a well characterized marker of angiogenesis) for 1 hour at room temperature. After washing the membrane with PBS, they were transferred on the multisensitive phosphor screen (Perkin-Elmer, USA) and imaged with Cyclone Plus Storage Phosphor System (Perkin-Elmer, USA).
9
Radioactive [18F]FDDNP Binding in Pancreas
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The [18F]FDDNP was prepared on the basis of a previously published method 16 (link). The dewaxed and hydrated human pancreas tissue sections were incubated in [18F]FDDNP PBS solution (0.13 MBq/ml) with or without its nonradioactive reference compound (0.44 mg/ml) in the dark for 20 min. The sections were then washed with distilled water and air dried before exposure to a multisensitive phosphor screen (PerkinElmer; Waltham, Massachusetts, USA) at RT for 5 min. The phosphor screen was then scanned in a Typhoon 8600 phosphor imager (GE Healthcare; Pittsburgh, Pennsylvania, USA) at a resolution of 50 µm. The images were analyzed using OptiQuant (version 5.0; PerkinElmer) software.
10
Papio anubis Nasal Cavity Autoradiography
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A freshly dissected Papio anubis nasal cavity was placed in freshly made 4% PFA in PBS and stored at 4°C. Tissue from either side of the nasal septum was carefully detached from the septum using forceps and placed in 10 mM Tris, pH 7.5, at 4°C. The tissue was then submerged for 20 minutes in 10 mM Tris buffer containing nonradiolabeled GV1-57 (10 or 0 μM) and 5% DMSO. To these solutions was added [11C]GV1-57 (~200 nCi), before incubation at RT for 15 minutes. The septal tissue was washed quickly by submerging 5 times in 10 mM Tris buffer and then transferred to a conical vial containing fresh 10 mM Tris buffer for 10 minutes at RT. The tissue was removed, air-dried, and exposed to a multisensitive phosphor screen (PerkinElmer) for 1 hour and imaged with a Cyclone Plus Storage Phosphor system (PerkinElmer). Images were colored using the color scale option in OptiQuant software (PerkinElmer).
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