FSOCA-induced switchable footpad skin optical clearing window for blood flow and cell imaging in vivo.

Abstract

The mouse footpad for its feature of hairlessness provides an available window for imaging vascular and cellular structure and function in vivo. Unfortunately, the strong scattering of its skin limits the penetration of light and reduces the imaging contrast and depth. Herein, an innovative footpad skin optical clearing agent (FSOCA) was developed to make the footpad skin transparent quickly by topical application. The results demonstrate that FSOCA treatment not only allowed the cutaneous blood vessels and blood flow distribution to be monitored by laser speckle contrast imaging technique with higher contrast, but also permitted the fluorescent cells to be imaged by laser scanning confocal microscopy with higher fluorescence signal intensity and larger imaging depth. In addition, the physiological saline-treatment could make the footpad skin recover to the initial turbid status, and reclearing would not induce any adverse effects on the distributions and morphologies of blood vessels and cells, which demonstrated a safe and switchable window for biomedical imaging. This switchable footpad skin optical clearing window will be significant for studying blood flow dynamics and cellular immune function in vivo in some vascular and immunological diseases. Picture: Repeated cell imaging in vivo before (a) and after (b) FSOCA treatment. (c) Merged images of 4 h (cyan border) or 72 h (magenta border) over 0 h. (d) Zoom of ROI in 4 h (yellow rectangle) or 72 h (red rectangle).