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Seebest: A pH-Adjustable Tissue Clearing Solution that Preserves Lipid Ultrastructures

Visualizing biological events and states to resolve biological questions is challenging. Tissue clearing permits three-dimensional multicolor imaging. Here, we describe a pH-adjustable tissue clearing solution, Seebest (SEE Biological Events and States in Tissues), which preserves lipid ultrastructures at an electron microscopy level. Adoption of polyethyleneimine was required for a wide pH range adjustment of the tissue clearing solution. The combination of polyethyleneimine and urea had a good tissue clearing ability for multiple tissues within several hours. Blood vessels stained by lipophilic carbocyanine dyes were deeply visible using the solution. Adjusting the pH of the solution was important to maximize the fluorescent intensity and suppress dye leakage during tissue clearing. The spatial distribution of doxorubicin and oxidative stress were observable using the solution. Hence, the Seebest solution provides pH-adjustable, rapid, sufficient tissue clearing while preserving lipid ultrastructures, which is suitable for multicolor deep imaging.