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Laser speckle imaging of blood flow beneath static scattering media

Abstract

Laser speckle imaging (LSI) is a wide-field optical imaging technique that provides information about the movement of scattering particles in biological samples. LSI is used to create maps of relative blood flow and perfusion in samples such as the skin, brain, teeth, gingiva, and other biological tissues. The presence of static, or non-moving, optical scatterers affects the ability of LSI to provide true quantitative and spatially resolved measurements of blood flow. With in vitro experiments using tissue-simulating phantoms, we determined that temporal analysis of raw speckle image sequences improved the quantitative accuracy of LSI to measure flow beneath a static scattering layer. We then applied the temporal algorithm to assess the potential of LSI to monitor oral health. We designed and tested two generations of miniature LSI devices to measure flow in the pulpal chamber of teeth and in the gingiva. Our preliminary clinical pilot data indicated that speckle contrast may correlate with gingival health.

To improve visualization of subsurface blood vessels, we developed a technique called photothermal LSI. We applied a short pulse of laser energy to selectively perturb the motion of red blood cells, increasing the signal from vasculature relative to the surroundings. To study the spectral and depth dependence of laser speckle contrast, we developed a Monte Carlo model of light and momentum transport to simulate speckle contrast. With an increase in the thickness of the overlying static-scattering layer, we observed a quadratic decrease in the quantity of dynamically scattered light collected by the detector. We next applied the model to study multi-exposure speckle imaging (MESI), a method that purportedly improves quantitative accuracy of subsurface blood flow measurements. We unexpectedly determined that MESI faced similar depth limitations as conventional LSI, findings that were supported by in vitro experimental data. Finally, we used the model to study the effects of epidermal melanin absorption on LSI, and demonstrated that speckle contrast is less sensitive to varying melanin content than reflectance. We then proposed a two-wavelength measurement protocol that may enable melanin-independent LSI measurements of blood flow in patients with varying skin types. In conclusion, through in vitro and in silico experiments, we were able to further the understanding of the depth dependent origins of laser speckle contrast as well as the inherent limitations of this technology.

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