Abstract
Tissue-clearing techniques have received great attention for volume imaging and for the potential to be applied in optical diagnosis. In principle, tissue clearing is achieved by reducing light scattering through a combination of lipid removal, size change, and matching of the refractive index (RI) between the imaging solution and the tissue. However, the contributions of these major factors in tissue clearing have not been systematically evaluated yet. In this study, we experimentally measured and mathematically calculated the contribution of these factors to the clearing of four organs (brain, liver, kidney, and lung). We found that these factors differentially influence the maximal clearing efficacy of tissues and the diffusivity of materials inside the tissue. We propose that these physical properties of organs can be utilized for the quality control (Q/C) process during tissue clearing, as well as for the monitoring of the pathological changes of tissues.
Highlights
Understanding the architecture of an organ or the body as a whole, using cellular resolution, is one of the ultimate goals in biology
We comprehensively analyzed the factors affecting tissue clearing, and we propose novel methods to evaluate the status of tissue clearing, which can be used as markers for tissue injury
Optical tissue clearing is achieved by the combination of 3 major factors: lipid removal, size expansion, and refractive index (RI) adjustment
Summary
Understanding the architecture of an organ or the body as a whole, using cellular resolution, is one of the ultimate goals in biology. The clearing-related properties of organs have not yet been comprehensively explored, which makes choosing the best clearing methods for the desired organs difficult. To this end, we developed assay systems to analyze the differential clearing properties of organs/tissues and to evaluate the quality of tissue clearing based on the changes in their macromolecule components and transparency. We developed assay systems to analyze the differential clearing properties of organs/tissues and to evaluate the quality of tissue clearing based on the changes in their macromolecule components and transparency This battery of assays allows for the systematic monitoring of the clearing processes, which can be used as quality control methods for the standardization of the clearing process. Our protocols can be used to verify the pathological changes of tissues, which will be a foundation for the label-free diagnosis of tissues based on optical clearing methods
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