Highway-rail grade crossing (HRGC) accidents pose a serious risk of safety to highway users, including pedestrians trying to cross HRGCs. A significant increase in the number of HRGC accidents globally calls for greater research efforts, which are not limited to the analysis of accidents at HRGCs but also understanding user perception, driver behavior, potential conflicting areas at crossings, effectiveness of countermeasures and user perception towards them. HRGC safety is one of the priority areas in the State of Florida, since the state HRGCs experienced a total of 429 injuries and 146 fatalities between 2010 and 2019 with a significant increase in HRGC accidents over the last years. The present study aims to conduct a comprehensive analysis of the HRGCs that experienced accidents in Florida over the last years. The databases maintained by the Federal Rail Administration (FRA) are used to gather the relevant information for a total of 578 crossings that experienced at least one accident from 2010 to 2019. In contrast with many of the previous efforts, this study investigates a wide range of various factors, including physical and operational characteristics of crossings, vehicle and train characteristics, spatial characteristics, temporal and environmental characteristics, driver actions and related characteristics, and other relevant information. The outcomes of this research will help better understanding the major causes behind accidents at the HRGCs in the State of Florida in a holistic way by considering a variety of relevant factors, which will assist the appropriate stakeholders with implementation of safety improvement projects across the state.