Experimental neurosensory retinal detachment was induced in mice as described previously7 (link). Briefly, animals were anesthetized and a sclerotomy was performed using a 25G micro-vitrealretinal blade. A 35G beveled cannula (World Precision Instruments; Sarasota, FL, USA, Cat# NF35BV-2), attached to a NanoFil-100 syringe (World Precision Instruments, Cat# NANOFIL-100) was used to inject 2–3 µL of Healon (1% Hyaluronic Acid) (Abbott Medical Optics; Santa Ana, CA, USA, Cat# 05047450842) between the photoreceptor and RPE layers. Care was taken to detach approximately half of the retina in each animal. Only the sclerotomy was performed on the fellow eye as a control. After 3 days, animals were sacrificed and whole eyes were extracted and embedded in paraffin for sectioning as described above. TUNEL staining was performed using the DeadEnd™ Fluorometric TUNEL System (Promega Cat# G3250) and sections were counterstained using ProLong Gold Mountant with DAPI. Stained sections were imaged with a Leica DM6000 microscope using a 40X objective. TUNEL positive cells were manually counted across the detached portion of the retina. Counts were normalized to the total number of nuclei in the outer nuclear layer in the detached region, counted manually or with an automated cell counting macro using ImageJ7 (link),56 (link).
Nanofil 100 syringe
Manufactured by World Precision Instruments
Sourced in United States
The NanoFil-100 is a precision syringe designed for accurate and controlled fluid dispensing. It features a glass barrel and stainless-steel plunger for durability and a minimal dead volume. The syringe has a volume capacity of 100 microliters and is suitable for a wide range of laboratory applications requiring precise liquid handling.
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Lab products found in correlation
3 protocols using nanofil 100 syringe
1
Retinal Detachment Model in Mice
2
Intravitreal Injections in Ocular Research
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Injections were delivered using a NanoFil-100 syringe with a 26G needle (World Precision Instruments, Sarasota, FL) under isoflurane (0.8% in O2; IsoThesia; Vetus Animal Health, Rockville Center, NY) inhalation anesthesia at a flow rate of 0.4 L/min using an Isoflurane Anesthesia machine for veterinary use only (Ohmeda Anesthesia Service and Equipment, Inc, Atlanta, GA). Following removal of the occluder, the sclera was exposed by retracting the eyelids with a handmade ocular speculum and injections were delivered through the sclera at the superior margin of the globe, just outside of the scleral ossicles, after cleaning the eyelids and surround area with 70% alcohol. Injections consisted of L-NAME (Sigma Chemical Co, St. Louis, MO) (a 30 μl injection containing 16.2 μmol of L-NAME in 0.9% saline), 30 μl of 0.9% saline (vehicle for L-NAME) (Nickla and Wildsoet, 2004 (link)), atropine sulfate (Sigma Chemical Co) (a 20 μl injection containing 240 nmol of atropine sulfate in phosphate-buffered saline, PBS), and 20 μl of PBS (vehicle for atropine sulfate) (Carr and Stell, 2016 (link)). The needle remained in place for 15 s before slowly withdrawing it from the eye and an ophthalmic antibiotic ointment (Vetropolycin, Pharmaderm, Melvill, NY) was applied to the eye. In some cases, the occluders were replaced prior to awakening from the anesthesia.
3
Intracranial Injection of Glioblastoma Cells in Nude Mice
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Five-week-old male nude mice (BioLASCO) were intracranially injected with 5 × 10 5 U-87 MG/Luc cells in 5 ml of growth medium. The tumor cell injection was performed at the position 0.5 mm to the right of the bregma, 2.5 mm posterior to the bregma, and 4 mm below the skull surface. Control or IGFBP3 siRNAs of 1.5 mg/mouse were mixed with in vivo jetPEI transfection reagent (Polyplustransfection) according to the manufacturer's instruction and infused into tumors using the convection-enhanced delivery (CED) method. Briefly, mice were anesthetized with inhalational anesthetic isoflurane (Forane, AbbVie Limited) and placed on a mouse stereotaxic instrument with continuous supplement of isoflurane. CED procedures were performed with a UMP3 microinjection system (UMC4 pump controller and UMP3 pump, World Precision Instruments) and a 100-ml NANOFIL-100 syringe (World Precision Instruments) with a 26G needle at an infusion rate of 1 ml/min. After infusion of 5 ml siRNA and in vivo jetPEI mixture, the cannula was left in the tissue for 10 minutes to prevent reflux. The weights of the mice and tumor sizes, determined by the Xenogen IVIS 100 In Vivo Imaging System, were measured twice a week. Mice were euthanized when they lost 20% weight or became weak and unable to perform normal living functions. The brains of mice were excised for further analyses.
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