© 2015 John Wiley & Sons Ltd. Objective: Gradual local heating of the skin induces a largely NO-mediated vasodilatation. However, use of this assessment of microvascular health is limited because little is known about its reproducibility. Methods: Healthy volunteers (n = 9) reported twice to the laboratory. CVC, derived from laser Doppler flux and mean arterial pressure, was examined in response to a standardized local heating protocol (0.5°C per 150 second from 33°C to 42°C, followed by 20 minutes at 44°C). Skin responses were examined at two locations on the forearm (between-site). Heating was repeated after a break of 24-72 hours (between-day). Reproducibility of skin responses at 33-42°C is presented for absolute CVC and relative CVC responses corrected for maximal CVC at 44°C (%CVCmax). Results: Between-day reproducibility of baseline CVC and %CVCmax for both sites was relatively poor (22-30%). At 42°C, CVC and %CVCmax responses showed less variation (9-19%), whilst absolute CVC responses at 44°C were 14-17%. Between-day variation for %CVCmax increased when using data from site 1 on day 1, but site 2 on the subsequent day (25%). Conclusion: Day-to-day reproducibility of baseline laser Doppler-derived skin perfusion responses is poor, but acceptable when absolute and relative skin perfusion to a local gradual heating protocol is utilized and site-to-site variation is minimized.