With the advent of fluorescent protein (FP) labeling, researchers have been interested in visualizing the expression of endogenous fluorescence within tissues and the visualization of these biomarkers in 3D has been of particular interest in various fields. It has been generally observed that non-aqueous clearing agents (BABB, DBE) and amphipathic clearing agents (Visikol HISTO) will quench FP over time. This creates problems for FP imaging as a tissue’s FP might be quenched prior to the tissue becoming transparent enough to visualize. While researchers can get around this problem by simply doing anti-FP immunolabeling, many researchers want a simple approach to clear tissue to look at FP.
To solve these problems, researchers have developed numerous aqueous clearing agents to preserve FP and one of those clearing agents is SeeDB which is a combination of fructose (80.2% wt/wt) in water and α-thiogylcerol (0.5%) at a refractive index of 1.48 ,. SeeDB is able to quickly render tissues transparent without causing swelling, but is not compatible with immunolabeling .