Background and Objective: Although cryogen spray cooling (CSC) is used to minimize the risk of epidermal damage during laser dermatologic surgery, concern has been expressed that CSC may induce cryo-injury. The objective of this study is to measure temperature variations at the epidermal-dermal junction in ex vivo human skin during three clinically relevant multiple cryogen spurt-laser pulse sequences (MCS-LPS). Study Design/Materials and Methods: The epidermis of ex vivo human skin was separated from the dermis and a thin-foil thermocouple (13 mm thickness) was inserted between the two layers. Thermocouple depth and epider-mal thickness were measured using optical coherence tomography (OCT). Skin specimens were preheated to 308C before the MCS-LPS were initiated. Three MCS-LPS patterns, with total cryogen spray times of 38, 30, and 25 milliseconds respectively, were applied to the specimens in combination with laser fluences of 10 and 14 J/cm 2 , while the thermocouple recorded the temperature changes at the epidermal-dermal junction. Results: The thermocouple effectively recorded fast temperature changes during three MCS-LPS patterns. The lowest temperatures measured corresponded to the sequences with longer pre-cooling cryogen spurts. No sub-zero temperatures were measured for any of the MCS-LPS patterns under study. Conclusions: The three clinically relevant MCS-LPS patterns evaluated in this study do not cause sub-zero temperatures in ex vivo human skin at the epidermal-dermal junction and, therefore, are unlikely to cause significant cryogen induced epidermal injury.