Abstract
Biological tissue transparency combined with light-sheet fluorescence microscopy is a useful method for studying the neural structure of biological tissues. The development of light-sheet fluorescence microscopy also promotes progress in biological tissue clearing methods. The current clarifying methods mostly use liquid reagent to denature protein or remove lipids first, to eliminate or reduce the scattering index or refractive index of the biological tissue. However, denaturing protein and removing lipids require complex procedures or an extended time period. Therefore, here we have developed acrylate resin with a high refractive index, which causes clearing of biological tissue directly after polymerization. This method can improve endogenous fluorescence retention by adjusting the pH value of the resin monomer.
Highlights
Biological tissue transparency combined with light-sheet fluorescence microscopy is a useful method for studying the neural structure of biological tissues
We investigated the effect of differences in the pH value of monomers on green fluorescent protein (GFP) fluorescence,[23] and a transparent GFP mouse brain block imaged via light-sheet fluorescence microscopy
The main effect of 2-phenoxyethyl acrylate is to increase the refractive index after polymerization, and the crosslinking agent triethylene glycol dimethacrylate causes the resin to form a crosslinked network structure that improves the hardness of the resin
Summary
Biological tissue transparency combined with light-sheet fluorescence microscopy is a useful method for studying the neural structure of biological tissues. We developed an acrylic resin with a high refractive index resin (termed HRAR) that clears biological tissue directly after embedding. The refractive index is increased when the liquid monomer polymerizes into solid resin, which matches the refractive index of the dehydrated biological tissue perfectly, to achieve transparency.
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