Laser Speckle Contrast Imaging (LSCI) is a wide field of view, non scanning optical technique for observing blood flow. Speckles are produced when coherent light scattered back from biological tissue is diffracted through the limiting aperture of focusing optics. Mobile scatterers cause the speckle pattern to blur; a model can be constructed by inversely relating the degree of blur, termed speckle contrast to the scatterer speed. In tissue, red blood cells are the main source of moving scatterers. Therefore, blood flow acts as a virtual contrast agent, outlining blood vessels. The spatial resolution (~10 μm) and temporal resolution (10 ms to 10 s) of LSCI can be tailored to the application. Restricted by the penetration depth of light, LSCI can only visualize superficial blood flow. Additionally, due to its non scanning nature, LSCI is unable to provide depth resolved images. The simple setup and non-dependence on exogenous contrast agents have made LSCI a popular tool for studying vascular structure and blood flow dynamics. We discuss the theory and practice of LSCI and critically analyze its merit in major areas of application such as retinal imaging, imaging of skin perfusion as well as imaging of neurophysiology.

Laser speckle contrast imaging (LSCI) is a high-resolution and high contrast optical imaging technique often used to characterize hemodynamic changes in short-term physiological experiments. In this study, we demonstrate the utility of LSCI for characterizing microvascular remodeling and hemodynamic changes during wound healing angiogenesis in vivo. A 2 mm diameter hole was made in the mouse ear and the periphery of the wound imaged in vivo using LSCI over 12 days. We were able to visualize and quantify the vascular and perfusion changes that accompanied wound healing in the microenvironment proximal to the wound, and validated these changes with histology. We found that consistent with the stages of wound healing, microvessel density increased during the initial inflammatory phase (i.e., day 0–3), stayed elevated through the tissue formation phase (i.e., until day 7) and returned to baseline during the tissue remodeling phase (i.e., by day 12). Concomitant ‘‘wide area mapping’’ of blood flow revealed that tissue perfusion in the wound periphery initially decreased, gradually increased from day 3–7, and subsided as healing completed. Interestingly, some regions exhibited a reestablishment of tissue perfusion approximately 6 days earlier than the ∼ 18 days usually reported for the long term remodeling phase. The results from this study demonstrate that LSCI is an ideal platform for elucidating in vivo changes in microvascular hemodynamics and angiogenesis, and has the potential to offer invaluable insights in a range of disease models involving abnormal hemodynamics, such as diabetes and tumors.

No abstract