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Antonina I Volikova1, Barry J Marshall1, J M A Yin2, Robert Goodwin2, Peter Ee-Pan Chow1, Michael J Wise1,3 1Marshall Centre for Infectious Disease Research and Training, School of Biological Sciences, University of Western Australia, Perth, Australia; 2Urology Department, Sir Charles Gairdner Hospital, Perth, Australia; 3Department of Computer Science and Software Engineering, School of Physics, Mathematics and Computing, University of Western Australia, Perth, AustraliaCorrespondence: Antonina I VolikovaUniversity of Western Australia, 35, Stirling Highway, Crawley WA 6009, AustraliaTel +61 845 015 4041Email Antonina.volikova@uwa.edu.auAim: The aim of this study was to apply and evaluate three ultrasound methods to measure the bladder wall in a healthy population using high-resolution applications and to establish reference points and baselines for future research into lower urinary tract diseases, specifically to understand how lower urinary tract disorders affect the bladder wall and to find objective, non-invasive diagnostic tests. Methods: The study was conducted on 116 healthy volunteers aged 19–79 years old with approximately 10 participants in each decade group. Results: The following bladder parameters were recorded and measured using a GE LOGIQ E9 XDclear 2.0 ultrasound machine (GE Healthcare, Wauwatosa, WI, USA): Full bladder wall thickness (BWT) and each of three bladder wall layers thickness (BWLT) – serosa, detrusor and mucosa;Shear Wave Velocity (SWV) in m/s, using 2D Shear Wave Elastography (2D-SWE); andBladder wall blood circulation (Resistive Index, RI), using Duplex Doppler ultrasound. All of the above measurements were recorded at three different urine filling volumes: V0 (20–50 mL), V2 (180–200 mL) and V4 (380–400 mL) with ten repeats for each measured parameter. As expected, BWT and BWLT correlated inversely with increasing bladder volume.While there are no large differences in the healthy bladders of men compared with women, or with age, some small, but statistically significant, differences revealed. BWT at V0 is greater in men, as is the detrusor thickness at VO, but there are no differences at other volumes or for other layers. There is a small, but statistically significant thickening ofBWTand detrusor layer and increase in SWVwith age in men at V0. SWE showed increase in SWVmeasured at 400 mL bladder volume across all gender and age groups. There was no change in bladder wall vessels RI with age, between gender groups or increasing bladder volume. Conclusion: We used three ultrasound applications to obtain bladder wall reference data in healthy individuals and investigated the relationships between BWT, BWLT, SWV, RI and gender, age at three bladder volumes, for further studies into identifying and diagnosing different urinary bladder disorders. With further research, ultrasound could be used as a diagnostic test to differentiate bladder pathology in clinical practice. Keywords: urinary bladder, healthy subjects, reference data, high-frequency ultrasound, duplex doppler, shear wave elastography