While efforts are being made to reduce opioid overdose fatalities, there is a need to equip individuals to act in overdose emergencies in an effort to slow the increasing rates of preventable opioid-related deaths in the United States. This study sought to determine whether education on naloxone administration would increase confidence in individuals utilizing harm reduction services to intervene in the presence of an opioid overdose.  Methods: A cross-sectional study was conducted that included adult participants attending Challenges, Inc., a mobile harm reduction service site. Participants completed a baseline survey assessing their level of comfort and experience with naloxone administration. An optional standardized educational session focusing on proper naloxone use followed, and all participants were offered free naloxone. Participants completed a post-survey reassessing their confidence and willingness to administer naloxone.  Results: Of the 100 participants, 75% reported witnessing at least one opioid overdose, with the majority of those seeing 10 or more opioid overdoses. Additionally, 58% of respondents had previously administered naloxone treatment, on average between one and four times. A significant increase was found in participants' confidence level of administering naloxone following the educational session (p < 0.0001). When asked about the likelihood of intervening when witnessing an opioid overdose, 96% agreed or strongly agreed that they would administer naloxone treatment.  Conclusion: A single education session increased the confidence, willingness, and comfort of individuals in administering naloxone in the presence of an opioid overdose. Ultimately, improving naloxone education and access could lead to a decrease in morbidity and mortality in association with opioid-related overdoses.