Multi-exposure Speckle Imaging for Quantitative Evaluation of Cortical Blood Flow.

Laser speckle contrast imaging (LSCI) is a label-free optical imaging technique that can quantify flow dynamics across an entire image. Multi-exposure speckle imaging (MESI) is an extension of LSCI that allows for reproducible and quantifiable measurements of flow. MESI has the potential to provide quantitative cerebral blood flow information in both preclinical and clinical applications; in fact, MESI can be extended to resolve the flow dynamics in any exposed tissue. A MESI system can be divided into three primary components: (i) the illumination optics, consisting of the optical source and a method of modulating and gating the illumination intensity; (ii) the collection optics, consisting of a high-speed camera that can be triggered and gated to match the pulsed illumination; and finally (iii) post-processing hardware and software to extract the flow information from the recorded raw intensity images. In the following protocol, we offer a guide to design, operate, and test a MESI system.