Purpose: Lesbian women are more likely to experience negative physical and mental health outcomes compared with heterosexual women, although most research on this population has relied on small convenience samples. This study compared health behaviors and health care utilization among lesbian women living in the South to representative subsamples of women from the general population. Methods: We conducted a Web-based survey of 1,141 self-identified lesbian women aged 19 and older living in the South. We compared descriptive results from our study to the U.S. Centers for Disease Control and Prevention's Behavioral Risk Factor Surveillance System. We also calculated adjusted logistic regression models to further explore correlates of Southern lesbian women's physical and mental health. Main Findings: Southern lesbians were less likely to be in poor physical health, but more likely to experience recent depression than women in the general population. Lesbians reported more risky health behaviors relative to other Southern and non-Southern women. Southern lesbians and other Southern women experienced similar barriers to routine health care, including lack of health insurance, not having a regular provider, and having to forgo care owing to cost. Many of these health behaviors and barriers to care were associated with poor physical and mental health among Southern lesbians. Conclusion: Southern lesbians' patterns of health behaviors and utilization of care may place them at increased risk for negative health outcomes relative to the general population. © 2010 Jacobs Institute of Women's Health.