Previous research suggests that individuals' sexual orientation may be affected by developmental instability (DI) induced by exposure to prenatal stresses. We tested this relationship using fluctuating asymmetry (FA), the small random deviations from symmetry that arise in otherwise bilaterally symmetrical traits as a consequence of developmental noise and developmental instability. Differences among individuals reflect variation in their exposure to and ability to accommodate for stresses experienced during development as well as to developmental noise that arises due to cellular stocasticity. FA measurements for 156 heterosexual and 132 homosexual men and women participants provided strong support for the developmental instability hypothesis: FA was significantly higher in both male and female homosexuals (men: four of seven bilateral traits and composite FA values (cFA); women: five of seven bilateral traits and composite FA values). Although finger-length ratios (FLRs), an indirect marker for prenatal hormones, were sex-atypical (e.g., feminized) for homosexual men, we failed to detect any relationship between FA levels and 2D:4D finger-length ratios (FLRs). Hence, although elevated levels of developmental stress appear to be linked to shifts in sexual orientation, the underlying mechanism does not seem to be connected to sex-atypical prenatal hormones. Additional analyses with sex atypical individuals are needed to confirm this.