Laser speckle imaging modality is one of widely used methods to evaluate blood flow because of its simplicity. However, laser speckle image has a limitation in the evaluation of subcutaneous blood flow due to its low contrast perfusion image. Various methods have been tried to enhance the perfusion image contrast. Such methods presented positive results in some degree. However, it could not be fundamental solutions due to low penetration depth of lasers restricted by optical tissue scattering property. This study suggests a method to enhance the perfusion image contrast of laser speckle imaging modality by increasing the penetration depth of lasers. An optical clearing agent (glycerol) was topically applied on skin treated with micro-needle roller in order to reduce the time period of optical tissue clearing and therefore, enhance the penetration depth of laser. In this study, we investigated the effect of glycerol and micro-needling methods in the contrast enhancement of laser speckle perfusion skin image and presented the results of in-vitro and in-vivo animal experiment.