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26 protocols using fluorinert

1

Ex Vivo MRI Analysis of Mouse Brain

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A small group of mice were anesthetized at PND19 using 4% isofluorane in air and sacrificed by decapitation. The entire heads were placed in tubes containing a cold solution of 4% Paraformaldehyde (PFA) and 8 mM Gadolium (Dotarem 0.5 mmol/ml, Guerbet). The tubes were then left at 4°C for 6–12 weeks for the contrast agent to fully diffuse and stabilize in tissue prior to ex-vivo imaging.
Magnetic Resonance Imaging (MRI) was performed on a 3T preclinical horizontal bore scanner (MR Solutions, Guilford, UK), equipped with a quadrature volume coil designed for rodent head imaging, with a 17 cm horizontal bore. On analysis day, the heads were placed in a custom-made 3D holder filled with fluorinert (3M, MN, USA) and anatomical and structural MRI sequences were acquired. Three dimensional Fast Spin Echo (FSE3D) anatomical series were used to calculate the volumes of brain sub-regions. Spin Echo Diffusion Tensor Imaging (SE-DTI) was used to calculate the diffusions parameters as described below. The details of each sequence have been reported in the Supplementary Data 6. The 3D Brain Atlas (Invicro's VivoQuant™) provided 14 sub-regions and the analysis of the changes in brain volume or in the diffusion parameters was performed with Matlab 2020b (MathWorks, MA, USA). The atlas provided 14 sub-regions and the analysis was completed for all sub-regions.
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2

Multiparametric MRI Protocol for Prostate Imaging

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All MRI studies were performed using a combination of an endorectal coil (BPX-30; Medrad, Pittsburgh, Pa) tuned to 127.8 MHz and a 16-channel cardiac coil (SENSE; Philips Medical Systems, Best, The Netherlands) on a 3-T magnet (Achieva; Philips Medical Systems, Best, The Netherlands) without previous bowel preparation. The endorectal coil was inserted using a semianesthetic gel (Lidocaine; AstraZeneca, Wilmington, Del) while the patient was in the left lateral decubitus position. The balloon surrounding the coil was distended with perfluorocarbon (3 mol/L-Fluorinert; 3M, St Paul, Minn) to a volume of approximately 45 mL to reduce susceptibility artifacts induced by air in the coil's balloon. The MRI protocol included triplanar T2W turbo spin echo, DW MRI (5 b values evenly spaced between 0 and 750 s/mm2), 3-dimensional MR spectroscopy, axial precontrast T1W, axial 3-dimensional T1W fast field echo dynamic contrast-enhanced MRI (DCE MRI) sequences, and their detailed sequence parameters are listed in Table 1. Dynamic contrast-enhanced MRI had 5.6-second temporal resolution, with Magnevist (Bayer, Whippany, NJ) injected after first 3 phases. An additional lower flip angle of 2° dataset was acquired before the DCE sequence for per-voxel T1 relaxation rate calculation to be used for quantitative DCE postprocessing.
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3

Multiparametric MRI Prostate Protocol

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The prostate mpMRI scans were acquired on a 3T scanner (Achieva 3.0T-TX, Philips Healthcare, Best, Netherlands) using an endorectal coil (BPX-30, Medrad, Pittsburgh, PA) filled with 45 mL fluorinert (3M, Maplewood, MN) and the anterior half of a 32-channel cardiac SENSE coil (InVivo, Gainesville, FL). Table 1 contains the sequences and MRI acquisition parameters used in this study.
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4

Infusion Protocols for Hippocampal and VTA Manipulation

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To help reduce any stress, all drugs were infused in the home cages. The stylets in the guide cannulae were replaced by a double infusion cannula (33 gauge, Plastics One) connected to two 5 μl microsyringes (WPI) in a microinfusion pump (Native Instruments) via flexible plastic tubing (C232CS, Plastics One) filled with Fluorinert (3M). The tips of infusion cannulae projected 0.5 mm below the tip of the guide cannulae. For intra-hippocampal microinjection, 0.5 μl of drug per cannula was infused at 0.2 μl min−1 (2.5 min). Infusion cannulas were left in place for a further 2.5 min before being replaced with stylets to aid drug absorption. For intra-VTA microinjection, 0.3 μl was injected at a rate of 0.3 μl min−1 (1 min) followed by 1-min of waiting. The mice were habituated to the experimental procedure of injection and to vehicle injection prior to the drug test in order to minimize novelty effect. Mice received drug injection 20 min (hippocampal microinfusions and i.p. injection of clonidine) or 3 min (VTA microinfusions) prior to the novelty exploration.
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5

Infusion Protocols for Hippocampal and VTA Manipulation

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To help reduce any stress, all drugs were infused in the home cages. The stylets in the guide cannulae were replaced by a double infusion cannula (33 gauge, Plastics One) connected to two 5 μl microsyringes (WPI) in a microinfusion pump (Native Instruments) via flexible plastic tubing (C232CS, Plastics One) filled with Fluorinert (3M). The tips of infusion cannulae projected 0.5 mm below the tip of the guide cannulae. For intra-hippocampal microinjection, 0.5 μl of drug per cannula was infused at 0.2 μl min−1 (2.5 min). Infusion cannulas were left in place for a further 2.5 min before being replaced with stylets to aid drug absorption. For intra-VTA microinjection, 0.3 μl was injected at a rate of 0.3 μl min−1 (1 min) followed by 1-min of waiting. The mice were habituated to the experimental procedure of injection and to vehicle injection prior to the drug test in order to minimize novelty effect. Mice received drug injection 20 min (hippocampal microinfusions and i.p. injection of clonidine) or 3 min (VTA microinfusions) prior to the novelty exploration.
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6

Endorectal MRI Imaging Protocol

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All MR imaging studies were performed using a combination of an endorectal coil (BPX-30; Medrad, Pittsburgh, PA, USA) tuned to 127.8 MHz and a 16-channel cardiac coil (SENSE; Philips Medical Systems, Best, the Netherlands) with a 3T magnet (Achieva: Philips Medical Systems, Best, the Netherlands), without prior bowel preparation. The endorectal coil was inserted using a semi-anesthetic gel (xylocaine, Lidocaine, Astra-Zeneca, Wilmington, DE, USA) while the patient was in the left lateral decubitus position. The balloon surrounding the coil was distended with 3-mol/L perfluorocarbon (Fluorinert; 3M, St Paul, MN, USA) to a volume of approximately 45mL, to reduce susceptibility artefacts induced by air in the coil’s balloon. The MR imaging protocol included triplanar T2-weighted turbo spin echo imaging, DW MRI imaging, axial three-dimensional fast-field-echo DCE MRI. Axial DCE images were obtained before, during, and after a single- dose of gadopentetate dimeglumine (Magnevist; Berlex, Wayne, NJ, USA), administered at a dose of 0.1 mmol per kilogram of body weight through a peripheral vein at a rate 3 mL/sec using a mechanical injector (Spectris MR injection System; Medrad, Pittsburgh, PA, USA). Sequence parameters were defined in previous studies [16 (link), 17 (link)].
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7

Ex Vivo Imaging of Perfused Brains

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Prior to ex vivo imaging, perfused brains were excised after day 7 post-MCAO and washed in PBS for 48 h. Brains were immersed in Fluorinert (3 M Corp, Minneapolis, Minnesota) for 24 h to minimize susceptibility artifacts [36 (link)] prior to imaging in a 15-mm NMR tube. Ex vivo images were acquired at (50-μm)3 isotropic resolution [TE = 7.5 ms, TR = 150 ms] using a 1H 3D GRE sequence at 11.75 T to assess cell dissociation from implanted aggregates.
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8

Multiparametric MRI Prostate Imaging Protocol

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MR images were acquired at 3-Tesla (Achieva 3.0T-TX, Phillips Healthcare, Best, Netherlands) using a combination of the anterior half of a 32-channel cardiac SENSE coil (InVivo, Gainesville, FL, USA) and an endorectal coil (BPX-30, Medrad, Pittsburgh, PA, USA) filled with 45 ml of fluorinert (3 M, Maplewood, MN, USA). The mpMRI protocol included acquisition of T2-weighted (T2W) sequences acquired separately in axial, coronal, sagittal plane, two diffusion-weighted imaging (DWI) sequence acquisitions consisting of one 5 b-value acquisition (0–750 s/mm2) for Apparent Diffusion Coefficient (ADC) estimation and one high b-value (2000 s/mm2), and Dynamic Contrast Enhanced (DCE) MRI. DCE MRI images were obtained in 5.6 s intervals following a single dose of gadopentetate dimeglumine 0.1 mmol/kg at 3 ml/s. Axial T1-weighted MR images of the abdomen were also obtained. Full acquisition parameters are listed in Supplementary Table 1.
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9

Microfluidic Dynamics of Galinstan and Liquids

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We introduced Galinstan (eutectic gallium indium stannum, Zairyo-ya.com) and liquid into a microchannel composed of PDMS (DuPont Toray Specialty Materials K.K.). We used water, vegetable oil (Kadoya Sesame Mill inc.), Fluorinert (3 M), and silicone oil (MOMENTIVE) as the liquids. One side of the through-hole of the PDMS microchannel was an inlet, and the other side was an outlet. We fabricated two kinds of channels, that is, a cylindrical channel and a square pole-shaped channel. The diameter of the cylindrical channel ϕ was 600 μm–1 mm, and the dimensions of the square pole channel were 500 μm × 500 μm. For observation of Galinstan in the microchannel, we used a microscope (DIGITAL MICROSCOPE VHX-500F, KEYENCE) and a transmission optical microscope (MX9430, MEJI TECHNO). All observation and experiments in this paper were performed at room temperature.
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10

Multiparametric MRI for Prostate Cancer

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MR images were acquired with a 3-T system (Achieva 3 T-TX, Philips Healthcare) and the combination of the anterior half of a 32-channel cardiac sensitivity-encoding coil (Invivo, Philips Healthcare) and an endorectal coil (BPX-30, Medrad) filled with 45 mL of perfluorocarbon-based fluid (Fluorinert, 3M). The mpMRI protocol included T2-weighted imaging (axial, coronal, sagittal), DWI (b = 2000 s/mm2), and dynamic contrast-enhanced MRI. DW images consisted of a high-b-value (2000 s/mm2) sequence and ADC map (from five evenly spaced b values of 0–750 s/mm2), which was the main sequence used for analysis. ADC maps were automatically calculated by the MRI unit software by means of a mono-exponential decay model fitted to data from images obtained at the five evenly spaced b values. The following model equation was used: S(b) = S(0) × exp(−b × ADC), where b is the gradient created by any one of the five b values used. The full mpMRI acquisition parameters are shown in Table 2.
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