For the plasma protein binding studies, the anesthetized animals (n = 3 for each tracer) were injected with [18F]F-DPA (27.0 ± 5.1 MBq) or [18F]DPA-714 (29.8 ± 3.4 MBq). After 15 min, the animals were sacrificed and the blood was collected into Li-heparin tubes and centrifuged (3000×g, 10 min). A 10-μl aliquot of the plasma was spotted onto a TLC plate; following this, the remaining plasma was filtered by ultrafiltration using Microsep 30K Omega (Pall Life Sciences, Mexico) with a cut-off of 30 kDa. A 10-μl aliquot of the resulting supernatant was also spotted onto the TLC plate. The plates were exposed onto an imaging plate (Fuji BAS Imaging Plate TR2025, Fuji Photo Film Co., Ltd., Tokyo, Japan) for approximately two half-lives of fluorine-18 (3.5 to 4 h). After the exposure, the imaging plates were scanned using a BAS-5000 reader (Fuji, Japan) with a resolution of 50 μm and the saved images were analyzed using TINA software (version 2.10 g, Raytest, Isotopenmessgeräte, GmbH, Straubenhardt, Germany).
Bas 5000 reader
Manufactured by Fujifilm
Sourced in Japan
The BAS-5000 reader is a laboratory instrument designed for analyzing and detecting radiation signals from samples. It is capable of reading and processing data from various types of radioisotope-labeled samples.
Automatically generated - may contain errors
Lab products found in correlation
Tina software, by Elysia raytest (2 mentions)
Cryostar nx50, by Thermo Fisher Scientific (2 mentions)
Cm3050, by Leica (1 mentions)
Wizard2 3, by PerkinElmer (1 mentions)
Superfrost ultra plus, by Thermo Fisher Scientific (1 mentions)
Eosin y, by Merck Group (1 mentions)
Superfrost plus, by Thermo Fisher Scientific (1 mentions)
Bas ms 2025, by Fujifilm (1 mentions)
Gill no 1, by Merck Group (1 mentions)
Sigmascan software, by IBM (1 mentions)
F254 tlc plate, by Merck Group (1 mentions)
Phosphor image plate, by Fujifilm (1 mentions)
7 protocols using bas 5000 reader
1
Plasma Protein Binding Assay for Radiotracers
2
Quantifying Regional Brain Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the ex vivo studies, the rats used for in vivo studies were sacrificed immediately after the 60 min PET scan. The brain was dissected and weighed, and the radioactivity was measured with a gamma counter (Wizard2 3”, PerkinElmer, Turku, Finland). The brain was then quickly frozen in isopentane (2-methylbutane; Sigma-Aldrich) on dry ice. Coronal brain sections of 20 µm were obtained using a cryostat (Leica CM3050S, Germany). The sections were mounted on a glass slide (Superfrost Ultra Plus, Thermo Fisher, USA). The slides were exposed to an image plate (Fuji BAS Imaging Plate TR2025, Fuji Photo Film Co., Ltd., Tokyo, Japan) for about two half-lives of the radioisotope in question.
After the exposure, the imaging plates were scanned with BAS-5000 reader (Fuji, Japan) with a resolution of 25 µm, and the saved images on the computer were analyzed by AIDA Image Analyzer 4.5 software (Raytest, Isotopenmessgeräte, Straubenhardt, Germany). The regions of interest were drawn on the LC, bed nucleus of the stria terminalis (BNST), neocortex (CTX), striatum (STR), and cerebellum (CB). The regions of interest (ROIs) were analyzed as photostimulated intensity/area (PSL/mm2) and presented as ratios relative to the CB. LC and BNST, CTX, and STR ratios to CB ratios were calculated in the adult and immature rats from the ARGs.
After the exposure, the imaging plates were scanned with BAS-5000 reader (Fuji, Japan) with a resolution of 25 µm, and the saved images on the computer were analyzed by AIDA Image Analyzer 4.5 software (Raytest, Isotopenmessgeräte, Straubenhardt, Germany). The regions of interest were drawn on the LC, bed nucleus of the stria terminalis (BNST), neocortex (CTX), striatum (STR), and cerebellum (CB). The regions of interest (ROIs) were analyzed as photostimulated intensity/area (PSL/mm2) and presented as ratios relative to the CB. LC and BNST, CTX, and STR ratios to CB ratios were calculated in the adult and immature rats from the ARGs.
3
Autoradiographic Mapping of CB2 Receptors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Autoradiography was performed on rat/mouse spleen tissue. Sections were prepared in 20 μm thickness using a cryostat (Cryo-Star NX 50; Thermo Scientific). The tissue slices were adsorbed to SuperFrost Plus (Menzel) slides and stored at −20°C until use. After thawing on ice for 10 min, sections were pre-incubated in the assay buffer containing 50 mM TRIS, 5 mM MgCl2, 2.5 mM EDTA, and 0.5% BSA (pH 7.4). Slices were dried and incubated in a humidified chamber with [11C]AAT-015 (5 nM) or [11C]AAT-778 (5 nM) in assay buffer for 15 min at room temperature. In order to test for specificity toward CB2, a solution containing both radiotracer and an excess of CB2 specific partial agonist GW405833 (10 μM) was prepared and added to the tissue. The tissue slices were washed twice with assay buffer (each 2 min) and twice with distilled water (each 5 s) on ice, air dried, and exposed to a phosphor imager plate for a period of 25 min. The plate was scanned using a BAS5000 reader (Fujifilm, Dielsdorf, CH).
4
Autoradiographic Quantification of Receptor Binding
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Coronal Wistar rat brain sections (10 μm) were cut on a cryostat (CryoStar NX50, Thermo Scientific, Waltham, MA, USA) and mounted on glass slides (Superfrost Plus, Thermo Scientific). Consecutive sections were thawed on ice for 10 min before pre-incubation in HEPES/BSA-buffer (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid/bovine serum albumin) (30 mM HEPES, 1.2 mM MgCl2, 110 mM NaCl, 2.5 mM CaCl2, 5 mM KCl, pH 7.4, 0.1% BSA) at 4 °C for another 10 min. Excess solution was carefully removed and tissue slices were incubated with 1 nM [18F]PSS232 supplemented with either vehicle (0.9% NaCl), 1 µM racemic ketamine, 1 µM S-ketamine, 1 µM ceftriaxone, 1 µM L-glutamate, 1 µM mGluR5 antagonist MMPEP or 1 µM mGluR5 antagonist ABP688. After incubation for 40 min at room temperature in a wet chamber, the slices were washed in ice cold HEPES/BSA-buffer for 5 min, twice in HEPES-buffer for 2 min each and finally dipped twice in distilled water. Dried slices were exposed to a phosphor imager plate (BAS-MS 2025, Fuji, Dielsdorf, Switzerland) for 15 min and the plate was scanned in a BAS-5000 reader (Fuji). A consecutive section was stained with hematoxylin (Gill No. 1, Sigma) and eosin (Eosin Y, Sigma) (HE) and digitalized by a slide scanner (Pannoramic 250, Sysmex, Horgen, Switzerland) to obtain brain morphology.
5
Aortic Autoradiography for Atherosclerosis Assessment
6
TLC Separation and Quantification of Plasma and Brain Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The blood samples were taken into Li-heparin tubes and centrifuged (3000×g, 10 min). The plasma was transferred to an Eppendorf tube and the plasma proteins were precipitated by addition of 1.5 parts of MeOH by volume. Following a further centrifugation (12,000×g, 4 min), 20 μl of the supernatant was spotted onto an aluminum-backed silica gel 60/Kieselguhr F254 TLC plate (EMD Millipore 1.05567.0001, Merck Millipore, Darmstadt, Germany).
Brain samples from the cortex were homogenized with approximately 200 μl of 1:1 v/v of MeOH and water; the mixture was transferred to an Eppendorf tube and centrifuged (12,000×g, 4 min). An aliquot (20 μl) of the supernatant was spotted onto the TLC plate.
The TLC plates were developed with a mixture of dichloromethane and methanol (9:1, v/v) for a distance of 6 cm. Following this, the plates were exposed onto an imaging plate (Fuji BAS Imaging Plate TR2025, Fuji Photo Film Co., Ltd., Tokyo, Japan) for approximately two half-lives of fluorine-18 (3.5 to 4 h). After the exposure, the imaging plates were scanned using a BAS-5000 reader (Fuji, Japan) with a resolution of 50 μm and the saved images were analyzed using TINA software (version 2.10 g, Raytest, Isotopenmessgeräte, GmbH, Straubenhardt, Germany).
Brain samples from the cortex were homogenized with approximately 200 μl of 1:1 v/v of MeOH and water; the mixture was transferred to an Eppendorf tube and centrifuged (12,000×g, 4 min). An aliquot (20 μl) of the supernatant was spotted onto the TLC plate.
The TLC plates were developed with a mixture of dichloromethane and methanol (9:1, v/v) for a distance of 6 cm. Following this, the plates were exposed onto an imaging plate (Fuji BAS Imaging Plate TR2025, Fuji Photo Film Co., Ltd., Tokyo, Japan) for approximately two half-lives of fluorine-18 (3.5 to 4 h). After the exposure, the imaging plates were scanned using a BAS-5000 reader (Fuji, Japan) with a resolution of 50 μm and the saved images were analyzed using TINA software (version 2.10 g, Raytest, Isotopenmessgeräte, GmbH, Straubenhardt, Germany).
7
In Vitro Autoradiography of [11C]7
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The in vitro autoradiography of [ 11 C]7 was carried out using an analogous procedure in the literature [10] (link). An aqueous buffer comprising 30 mM HEPES, 1.2 mM MgCl 2 , 110 mM NaCl, 5 mM KCl, 2.5 mM CaCl 2 , and 1% fatty acid free bovine serum albumin (pH ~7.4) was employed as incubation buffer. The brain sections were incubated with [ 11 C]7 (18 nM, 65.6 GBq/μmol) in a humidified chamber for 30 min. Blocking experiments on Wistar brain sections were carried out in the presence of 10 μM SAR127303 or 10 μM PF-06795071. Upon the completion of incubation, the slices were washed in the abovementioned incubation buffer (pH 7.4, 0 • C, 1 × 2 min), washing buffer comprising 30 mM HEPES, 1.2 mM MgCl 2 , 110 mM NaCl, 5 mM KCl, 2.5 mM CaCl 2 (pH 7.4, 0 • C, 2 × 2 min), followed by 2 quick dips in distilled water (0 • C, 2 × 5 s). After drying, the slices were attached to a phosphor image plate (Fuji, Dielsdorf, Switzerland) and the exposure lasted for 60 min. The film was scanned by BAS5000 reader (Fuji), and the autoradiogram was analyzed by AIDA 4.50.010 software (Raytest Isotopenmessgeräte GmbH, Straubenhardt, Germany).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!