It is now established that the bladder is not sterile; it contains communities of microbes (microbiota). While the healthy bladder microbiota have been defined using expanded quantitative urine culture (EQUC) and urines obtained by transurethral catheterization (TUC), longitudinal and population studies have not previously been possible. These studies cannot be done using TUC urines, as it would be impractical to catheterize participants daily. Instead, we must transition to using voided urines.In order to make this switch, we have addressed three main issues raised by using voided urine. First, we showed that EQUC out performs standard urine culture on voided urines in a clinically relevant population. Next, we defined the constituents of the urethral and peri-urethral microbiota in an attempt to deconstruct voided urine. Finally, we identified a cleaner catch method. Combined, these studies provide crucial insight for interpreting voided urine for urinary microbiome research