Purpose To compare retinal microvascular function in healthy individuals with and without a positive family history (FH) of cardiovascular disease (CVD). Methods Retinal vessel reactivity was assessed by means of dynamic retinal vessel analysis in 38 healthy subjects aged between 30 and 66 years with a positive FH of CVD and 37 age‐ and gender‐matched control subjects. Other assessments included blood pressure (BP) profiles, blood glucose and lipid metabolism markers, Framingham risk scores (FRS), carotid intima‐media thickness (c‐IMT) and brachial flow‐mediated dilation (FMD). Results Family history‐positive subjects showed decreased retinal arterial baseline diameter fluctuation, dilation amplitude, percent dilation, and overall constriction response slope (p = 0.001; p = 0.015; p = 0.001; and p < 0.001, respectively) and increased percent constriction (p = 0.008). On the venous side, baseline‐corrected flicker response and dilation response slope were decreased in the FH‐positive group (p = 0.009 and p = 0.010, respectively). There were no significant differences between groups in c‐IMT scores or FMD parameters (all p > 0.05). The arterial MC% correlated negatively with decreased high‐density lipoprotein cholesterol (r = −0.52, p = 0.002) in only FH‐positive group. Conclusion Although macrovascular function is preserved in individuals with FH positive for CVD but with low FRS, there are, however, functional impairments at the retinal microvascular level that correlate with established plasma markers for cardiovascular risk.