Needles were made from glass capillaries (25-μl Drummond Microcaps) using a needle puller (Narishige), and the tip was sharpened using a needle grinder (Narishige). The needle gauge was adjusted to 12–14 μm. A micromanipulator (Leica) equipped with a glass needle was used for transplantation.
Needle puller
Manufactured by Narishige
The Needle Puller is a device used to manually pull and remove pipette or microinjection needles from needle holders. It is designed to allow for precise and controlled needle extraction, preventing potential damage to the needle or the needle holder.
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Lab products found in correlation
2 protocols using needle puller
1
Glass Capillary Needle Fabrication
2
Fluorescent Lipophilic Dye Labeling
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Fluorescent lipophilic dyes DiI (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate, Molecular probes D-282) and DiA (4-(4-(dihexadecylamino)styryl)-N-methylpyridinium iodide, Molecular probes D-3883) were used at a stock concentration of 2.5 mg/mL in dimethylformamide (DMF). Glass capillary needles (FHC Cat. 30-30-0) with an O.D of 1.0 mm were pulled using a Narishige needle puller to create tapered ends. Capillaries were loaded with dye, mounted in a capillary holder and moved into position for injection using a Narishige micromanipulator mounted to a stereomicroscope. Labeling was performed with a 3-millisecond (msec) pulse from a PicoSpritzer model II (General Valve). Labeling was confirmed by fluorescent microscopy. After 48 hours of incubation at 38°C, embryos were removed from the egg, dissected in phosphate-buffered saline (PBS), and fixed in 4% paraformaldehyde (PFA) before imaging and/or tissue processing.
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