We used 0.5% solutions of a small molecular weight (3 kDa) Texas Red®-conjugated dextran (TR-d3, Thermo Fisher Scientific) and a large molecular weight (40 kDa) fluorescein isothiocyanate (FITC)-conjugated dextran (FITC-d40, Sigma-Aldrich) in a 1:1 ratio dissolved in the artificial CSF (aCSF, Tocris Bioscience, Bristol, UK) for the co-infusion experiment [32 (link)]. To prevent leakage of the CSF tracers, the cannula was tightly arranged in a series as follows: A Hamilton 700 series syringe with a 22-gauge needle (Sigma-Aldrich) was connected to PE50 and PE10 polyethylene tubing. The other end of the PE10 tubing was linked to a 28-gauge injection cannula inserted into the cisterna magna, through which the CSF tracers moved. The rats were initially induced with 3% isoflurane and then anesthetized intraperitoneally with a combination of Zoletil® 50 (30 mg/kg) and xylazine (10 mg/kg). Anesthetized rats were fixed in a stereotaxic frame while the atlanto-occipital membrane overlying the cisterna magna was surgically exposed. A total volume of 70 μL of the CSF tracers was infused at a rate of 1.6 μL/min using a syringe pump. After the infusion was finished, the needle was left in place for 30 min. Then, the anesthetized rats were transcardially perfused and fixed with 4% PFA. Fluorescence imaging was performed on 30 μm coronal sections of the brain tissue.
Hamilton 700 series syringe
Manufactured by Merck Group
Sourced in United States
The Hamilton 700 series syringe is a laboratory instrument designed for precise liquid handling. It features a durable glass syringe and a robust design to ensure accurate and reproducible results. The syringe is available in a range of volume capacities to accommodate various application requirements.
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2 protocols using hamilton 700 series syringe
1
Co-Infusion of CSF Tracers in Rats
2
Neonatal Mouse Brain Vector Injections
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Animal experiments reported in this study were conducted with C57/Bl6 mice bred and maintained in accordance to NIH guidelines as approved by the UNC IACUC. Pups at post-natal days 1–2 were rapidly anesthetized on ice for 2 min followed by stereotaxic i.c.v. injections. Specifically, vectors packaging different transgenes were injected at a dose of 4 × 109 vector genomes into the left lateral ventricle (total volume < 3 μL) using a Hamilton 700 series syringe with a 26s gauge needle (Sigma-Aldrich, St. Louis, MO, USA), attached to a KOPF-900 small-animal stereotaxic instrument (KOPF Instruments, Tujunga, CA, USA). All neonatal injections were performed 0.5 mm relative to the sagittal sinus, 2 mm rostral to transverse sinus, and 1.5 mm deep. Following vector administration, mice were revived under a heat lamp and rubbed in the bedding before being placed back with the dam. At 6 weeks after injection, mice were overdosed with tribromoethanol (Avertin) (0.2 mL/10 g of 1.25% solution) via the intraperitoneal route. This was followed by transcardial perfusions of 4% paraformaldehyde in PBS. The brain was harvested and postfixed in 4% paraformaldehyde for 24 hr. For this study, n = 3 mice were injected and processed for each construct.
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