We are developing a real-time infrared imaging technique, Enhanced Thermal Imaging(ETI), that can detect blood vessels embedded in tissue and assess tissue perfusion. ETI is a combination of standard thermal imaging (8 - 10 μm) and selective heating of blood relative to surrounding water-rich tissue using LED sources at low power. Blood absorbs strongly at 530 nm. Illumination of water-rich tissue and embedded blood vessels at this wavelength selectively increases the temperature of the blood vessels relative to the surrounding tissue causing the vessels to appear brighter in a thermal image. ETI does not require the use of injectable dyes and has a compact footprint allowing for use both during surgery and at the bedside. Previous studies using ETI were limited due to lengthy post-processing times re- quired to delineate vessels. The first study in this dissertation shows the real time capabilities of ETI in mapping vascular structures. Real time application of com- putational filters highlighting temporal and spatial changes reveal embedded blood vessels. The temporal filter averages the frames captured before LED illumination and then subtracts that average frame from each subsequent frame. The result is a 2-dimensional map showing how each pixel in the image changes temperature as the result of LED illumination. The spatial filter applies a 2-dimensional gradient function across each frame. This creates a 2-dimensional map showing the difference for each pixel from the neighboring pixels, thus revealing temperature boundaries. ETI was obtained of a model with simulated blood vessels and a porcine heart tissue model. In both sets of experiments the temporal and spatial filters outperformed standard thermal imaging. The temporal filter consistently increased the contrast between the vessel/tubing and the surrounding tissue. This increase in contrast al- iv lows for the temporal filter to detect and map the vessels more quickly and accurately than standard thermal imaging. The application of the spatial filter consistently de- lineated the vessel walls clearly and provided measurements of the vessel width. The spatial filter images have potential to highlight the presence of occlusions in vessels. In the second study presented in this dissertation, simulations of the illumination and heating of the blood vessels embedded in tissue were conducted to understand the effects of LED power and vessel depth on the ability of ETI to detect vessels. The simulations were performed with an open-source MATLAB integrated solver, MCmatlab. A tissue model with a volume of 1 cm3 consisting of air, epidermis, dermis, and a single blood vessel was simulated using several different LED illumination powers (0.1 - 1 W) and different vessel depths (1 - 3 mm). ETI can be used in two different modalities, direct and indirect illumination. Both modalities were simulated. In direct illumination the area of illumination is the same as the area of imaging. In indirect illumination, the region of interest for imaging is downstream (in direction of blood flow) from the site of illumination. The objective of the simulations was to characterize the limitations of ETI, optimize imaging parameters and understand the effects of vessel depth and width on imaging. The results of the simulations were compared to ex vivo tissue experiments of ETI using porcine skin, blood, dissected kidney vessels, and intact porcine hearts. In the third study presented in this dissertation, ETI was used to monitor the heal- ing of skin flaps in a murine model. ETI, fluorescent imaging and visual inspection were used to assess the viability of skin flaps in 15 mice over a 12 day study period. The results from these three different techniques were then compared. ETI plus the temporal derivative filter revealed persistent healing not seen using the other tech- niques. This study shows the potential of ETI to assess re-perfusion post-operatively with insights into non-superficial healing