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Introduction Squirting is the involuntary expulsion of fluid from the female urethra following stimulation of the anterior vaginal wall before or during orgasm. The mechanism underlying squirting has not been established. Purpose To elucidate the mechanism of squirting. Methods The subjects in the current study were women who were able to squirt. They were not sex workers. A urethral catheter was inserted before sexual stimulation and the bladder was emptied. Then, a mixture of indigo carmine (10 ml) and saline (40 ml) was injected into the bladder. Sexual stimulation was provided to facilitate squirting, which was videotaped and verified. The secretions were collected in sterile cups, and prostate specific antigen (PSA) and glucose levels were measured. Results Five women (2 in the 30s, 2 in the 40s, and 1 in the 50s) participated in this study. All women were able to squirt; three squirted only with manual sexual stimulation and two with penetrative sexual stimulation. The discharged fluid was blue in all cases, confirming the bladder as the source. The fluid was PSA-positive in four patients. Conclusions The main component of squirt fluid is urine, but may also contain fluid from Skene's glands (female prostate). This is the first report in which visualization of squirting was enhanced.