Purpose: Little is known about trans womens' experiences accessing gender-segregated health and housing services, particularly services for marginalized individuals living in poverty. As such, we conducted a qualitative investigation into experiences of accessing women-specific health and housing services among trans women and two-spirit persons in a downtown neighborhood of Vancouver, Canada. Methods: Between June 2012 and May 2013 interviews were conducted with 32 trans women and two-spirit individuals who had accessed women-specific health and/or housing services. Participants were recruited from four open prospective cohorts of sex workers and individuals who use drugs. Interview data were analyzed using a participatory analysis approach with two participants who were hired as research assistants. Results: Participants were generally able to access women-specific services in the neighborhood. However, there were reports of discrimination related to gender identity, discrimination based on gender expression (e.g., requirement of a feminine gender expression), and lack of staff intervention in harassment from other service users. Conclusion: Trans women and two-spirit persons in our study relied upon services for their health and safety and, therefore, exclusion from women-specific services had potentially severe adverse consequences such as homelessness and sexual violence. Recommendations to improve accessibility, including policy development and procedural recommendations, are put forth.