Methylene Blue Dye Injection in Mouse Embryonic Urinary Tract: A Method To Assess the Congenital Obstruction in the Urinary Tract

프로토콜

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board

1. Preparation of Methylene Blue Dye Solution

1. Measure 0.1 g methylene blue powder.
2. Dissolve the methylene blue in 10 mL normal saline or 1x phosphate-buffered saline (PBS) completely by vortexing.
3. Filter the 10 mg/mL methylene blue solution with a syringe filter (membrane pore size 0.45µm) to eliminate clogging during injection.
4. Assemble a sterile scalp vein set (27 G x 3/4″) with a 3 mL disposable syringe and fill the syr....

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결과

Figure 1. Examples of methylene blue dye injection into a normal urinary system. (A) A typical image of the urinary system from a perinatal embryo (E19.5) after removing excess tissue. Two kidneys, two ureters, and a bladder are seen with reproductive organs, here testes (B) An example of dye injection into a normal kidney. An injection point into the renal pelvis of the kidney (black arrow) can be seen. A weak dye color is visible in the ureter and the bladder ....

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자료

Name	Company	Catalog Number	Comments
Methylene blue	Sigma-Aldrich	M9140
Quality Biological Inc. NORMAL SALINE - 500ML	Fisherscientific	50-983-204
3 mL disposable syringe with BD Luer-Lok tip	BD	309657
Acrodisc Syringe Filters with Supor Membrane	Pall corporation	4614
Exel Scalp Vein (Butterfly) Sets; 27G x 3/4" 12"	EXELINT	26709
Delicate Operating Scissors	Roboz Surgical Instrument Co.	RS-6702
Micro Dissecting Forceps	Roboz Surgical Instrument Co.	RS-5135
Dumont Tweezers; Pattern #5	Roboz Surgical Instrument Co.	RS-5045
BD PrecisionGlide Needles 30G x 1/2"	BD	305106