This article details the procedure for emptying and washing a mouse's urinary bladder, which is essential for various experimental protocols. The method ensures sterile conditions and proper handling of the mouse during the process.
To empty and wash a mouse's urinary bladder, begin by taking an anesthetized female mouse in the supine position. Next, obtain an appropriately-sized catheter. Lubricate the tip of the catheter to reduce friction and ensure a smooth urethral insertion. Then, slowly and gently insert the lubricated catheter, passing it through the urethral opening, into the bladder.
Upon successful catheterization, the urine begins to drain out of the bladder into the catheter. Now, empty the urine-filled catheter. Using a pipette, add a suitable buffered saline into the outer end of the catheter while its opposite end is within the bladder. To the outer end of the catheter, then attach a syringe. Gently push the syringe to deliver the saline to the bladder.
Now, remove the syringe. Allow the saline, along with the residual urine, to drain out of the bladder through the catheter. Subsequently, vacate the residual urine mixed with saline from the catheter. Repeat the bladder washing with saline twice to ensure complete removal of the urine from the bladder. The prepped mouse is finally ready for further experimental procedures.
To begin this procedure, autoclave all surgical instruments and sterilize the workspace with 70% ethanol. Spray surgical instruments and the gloves with 70% ethanol frequently when performing the subsequent procedures to maintain sterile conditions.
To begin, anesthetize the mouse and shave the surgical area. To deplete the urine, apply lubricant to the catheter and ensure that its outer surface is fully covered. Next, insert the catheter into the urethral opening of the mouse and slowly push it until it reaches the bladder. Gently press the abdomen to help urine depletion.
Subsequently, discard the urine with a pipette into a waste beaker. Then, pipette 80 microliters of PBS into the outer end of the catheter with the other end still in the bladder. Next, carefully attach a 1-milliliter syringe to the catheter. Gently push the syringe to inject PBS into the bladder.
Leave some PBS in the catheter to avoid creating air bubbles in the bladder. Afterward, remove the syringe and wait for PBS to drain out of the bladder. Gently press the abdomen to evacuate PBS. Then, discard the PBS in the catheter with the pipette into the waste beaker. Repeat PBS washing for two more times.
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