All experiments were approved by the Institutional Animal Care and Use Committee of the Weizmann Institute of Science under protocol number 10790514–1. Fresh saccharomyces cerevisiae Baker’s yeast cells were dissolved in PBS in a 10 mm NMR tube, and left for ~72 hours prior to their MR investigation. Two mice were sacrificed by isoflurane overdose and their brains were fixed in formaline, and washed twice with PBS prior to their insertion to a 10 mm NMR tube filled with Fluorinert (Sigma-Aldrich, Rehovot, Israel). All specimens were left in the magnet for at least three hours prior to experiment commencement, to thermally equilibrate.
Fluorinert
Manufactured by Merck Group
Sourced in Portugal, United States
Fluorinert is a line of perfluorinated liquids developed by Merck Group. These liquids are designed for use in specialized laboratory equipment and industrial applications. They exhibit properties such as chemical inertness, low surface tension, and a wide liquid range, making them suitable for various testing and processing requirements.
Automatically generated - may contain errors
12 protocols using fluorinert
1
Yeast and Mouse Brain MRI Analysis
2
Transverse Relaxation Decay Analysis of Bacterial Nanocellulose
3
Postexcision Hippocampal Preparation for MRI
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Hippocampi were coronally cut postexcision, with the anterior (head) portion designated for clinical histopathological analyses and the posterior portion (body) for this study. The posterior portion was fixed immediately following surgery in formaldehyde (4%) for 48 hr before transfer and storage in phosphate buffered saline (PBS) at 4°C. Specimens (~20 × 12 × 12 mm) were prepared for MR imaging by immersion in proton‐free Fluorinert (Sigma‐Aldrich, St. Louis, MO) inside a syringe with a Luer cap for long duration immobilization, while avoiding the generation of air bubbles and dehydration.
4
Spinal Cord Extraction and NMR Imaging
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One rat spinal cord was extracted via transcardial perfusion with 4% Paraformaldehyde (PFA). After extraction, the spinal cord was immersed in a 4% PFA solution for 24 h, and then washed in a Phosphate-Buffered Saline (PBS) solution for 24 h. Two sections of cervical spinal cord were cut and placed separately in 5 mm NMR tubes filled with Fluorinert (Sigma Aldrich, Lisbon, PT). The samples were imaged on a 16.4 T Bruker Aeon Ascend scanner (Bruker, Karlsruhe, Germany) equipped with a 5 mm birdcage coil and gradients capable of producing up to 3 T/m in all directions.
5
High-Resolution Ex Vivo MRI of Brains
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All magnetic resonance imaging (MRI) was performed on a 7.0 T horizontal small bore magnet (Varian, Palo Alto, CA, USA) with a custom built 39 mm diameter head quadrature RF coil (David Herlihy, Imperial College London) linked to a Linux-based control console running VnmrJ acquisition software (v2.3, Varian, Palo Alto CA, USA). All acquisitions were performed at room temperature (21°C) on post-mortem samples with brains retained within the skull to avoid artifacts due to dissection or motion, as well as to avoid a tissue-air interface, which could affect cortical measurements. Prior to scanning, skulls were immersed in Fluorinert (Sigma) to provide a hydrogen-free background signal. Ex vivo scanning enables long scanning times that result in high spatial resolution images, which reduce potential confounding imaging artifacts, such as partial volume effects or head motion.
6
High-Field MRI of Fixed Brain Samples
7
High-Resolution 3D MRI of Intact Mouse Brain
8
Perfusion and MRI Preparation of Rat Brains
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Twenty-four hours after the last day of repeated inescapable stress or after daily handling in control animals, rats were anesthetized with sodium pentobarbital (100–150 mg kg−1, intraperitoneally (i.p.)), followed by intracardiac perfusion with physiological NaCl solution and 4% cold paraformaldehyde in 0.01 M phosphate-buffered saline (pH=7.4). After perfusion, the brain was harvested maintaining integrity and stored in 4% PFA in phosphate-buffered saline at 4 °C. Before MRI, the brains were washed into phosphate-buffered saline for 24 h to remove the fixation solution and then placed into a custom-built MRI-compatible tube. The tube was filled with Fluorinert, an MRI susceptibility-matching fluid (Sigma-Aldrich, St Louis, MO, USA).
9
MRI Phantom and Brain Sample Preparation
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Doped water phantoms were prepared by gradually adding copper sulfate to a 30/70% (volumetric) D2O/water, until a longitudinal relaxation time of approximately 200 ms was obtained. The solution was placed in a 5‐mm NMR tube, which was sealed and placed in a 10‐mm NMR tube filled with Fluorinert (Sigma Aldrich, Lisbon, Portugal). Brain samples (n = 3) were extracted from healthy male C57bl mice weighing approximately 25 g by standard intracardial paraformaldehyde perfusion, followed by 12 h in a 4% paraformaldehyde solution at 4 ºC, and placement in phosphate‐buffered saline at 4 ºC. The brains were then soaked in a solution of phosphate‐buffered saline and 0.5M gadoterate meglumine (Dotarem, Guerbet, Lisbon, Portugal) at a dilution of 1:200 (2.5 mM) for 12 h 41 , washed with phosphate‐buffered saline, and placed in a 10‐mm NMR tube filled with Fluorinert. All samples were allowed to equilibrate with the surrounding constant temperature of 37 ºC.
10
Spinal Cord Extraction and NMR Imaging
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