Background: Physical examination of potential organ and tissue donors is standard practice to mitigate risks and optimize outcomes for transplant recipients, but the content and process of the examination has not been investigated. The aim of this study was to determine current practice of performing a physical examination on potential organ and tissue donors in Australia. Methods: An online cross-sectional survey was circulated to all Australian Donor Coordinators (n = 125). Results: There were 75 responses (60% response rate) to the online survey. Respondents perform a mean 10.5 physical examinations per year. Inconsistencies were observed in the approach to the physical examination, inclusive of assessment techniques used to perform the examination such as palpation. Specific staff training and education to perform the examination was reportedly provided to 77% of respondents. There was less variation reported in examination findings classified as higher risk and escalation procedures with the 3 most common findings of injection sites / track marks (86%), suspicious moles (77%), and unexplained scarring (51%), and with 97% seeking a second opinion. Current and previously removed melanomas were the main examination findings that stopped a donation from proceeding, as reported to have occurred by 18 respondents. Conclusions: This study has identified variations in current physical examination practice and provided the evidence to pursue practice improvement. The inconsistencies can be partly attributed to discrepancies in training and education of staff and no standardized national guidelines to clearly outline expected practice.