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Bioimaging Core - Leuven
Several methods have been published describing different clearing protocols which can be divided into two large groups:
Organic solvent clearing protocols
The majority of these procedures have a simple two step mechanism in common:
Dehydration of the sample by removing water
Matching refractive indices
This clearing leads to very transparent samples and is perfectly suited for dense tissue like tumors, adult tissue, or highly myelinated brain. The majority of immuno-histochemical stainings are well conserved.
Water-based clearing protocols
The most common principle of water-based clearing protocols is by depolymerization. By splitting large structures like lipid bilayers into small micelles of different sizes, the opacity is remarkably reduced. Different clearing protocols vary in complexity and in degree of translucency that can be achieved. By depolymerization, the entire structure of a sample can be debilitated while the fluorescence of proteins like GFP is well preserved.