Title: Tissue Clearing With the PEG Associated Solvent System (PEGASOS)
Dian Jing, Texas A&M University
Shiwen Zhang, Texas A&M University
Wenjing Luo, Texas A&M University
Hu Zhao (Presenter)
Texas A&M University
Objectives: Tissue clearing technique enables visualization of opaque organ and tissue in 3-dimension by turning tissue transparent. Solvent-based tissue clearing methods yields better transparency than aqueous clearing methods, but also leads to significant loss of fluorescence. Here, we developed a polyethylene glycol (PEG) Associated Solvent System (PEGASOS), which efficiently renders both hard and soft tissue highly transparent within a short time
Methods: The PEGASOS method is composed of multiple treatment steps including fixation, decalcification, decolorization, delipidation, dehydration and final clearing. The entire process takes 7-14 days to complete depending on the type of tissue to be cleared. The reagent cost is inexpensive and the operation is easy to be reproduced in any regular lab.
Results: The PEGASOS method enables us to image the whole mouse brain with high resolution and to track individual neurons over a long distance within intact rodent brains. With the PEGASOS method, we were able to map connections between the peripheral and the central nervous system by imaging through the vertebrae. Importantly, we revealed the nerve distribution pattern within the long bone marrow space.
Conclusions: These results suggest that the PEGASOS method can be a powerful tool for biomedical research.