Abstract
The use of clearing agents has provided new insights in various fields of medical research (developmental biology, neurology) by enabling examination of tissue architecture in 3D. One of the challenges is that clearing agents induce tissue shrinkage and the shrinkage rates reported in the literature are incoherent. Here, we report that for a classical clearing agent, benzyl-alcohol benzyl-benzoate (BABB), the shrinkage decreases significantly with increasing sample size, and present an analytical formula describing this.
Highlights
Solvent-based tissue clearing is a widely used methodology to render an otherwise opaque sample optically transparent
Using tissue clearing in combination with various optical imaging techniques enables the study of various organs and organisms, such as mouse brains [1,2,3], larvae and spinal cords [4] as well as tumours including theirenvironment [5] such that the overall structure and interaction with the neighboring tissue structures can be studied and understood
We have shown that the tissue shrinkage using benzyl-alcohol benzyl-benzoate (BABB) clearing method is not fixed but depends on the sample size
Summary
Solvent-based tissue clearing is a widely used methodology to render an otherwise opaque sample optically transparent. Using tissue clearing in combination with various optical imaging techniques enables the study of various organs and organisms, such as mouse brains [1,2,3], larvae and spinal cords [4] as well as tumours including their (micro)environment [5] such that the overall structure and interaction with the neighboring tissue structures can be studied and understood. This has been important for the advancing understanding in neurology and oncology. A third category is hydrogel-based clearing methods, in which biomolecules are covalently
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