Evaluation of 2,2´-Thiodiethanol as a clearing agent for whole organ visualization in 3D microscopy

Optical tissue clearing has become a fundamental step in whole organ-volumetric analysis in 3D fluorescence microscopy. The need of developing new optical clearing agents that can render a whole mouse organ or human tissues transparent in an artifact-free way has led to the scientific community to invest great efforts in this field. As a result, many different clearing protocols have been developed in recent years, characterized by the quality of their interaction with the tissue in different aspects.

In this project we study optical clearing of mouse heart with 2,2´-Thiodiethanol (TDE), a new clearing agent recently introduced into the whole organ-optical clearing research field. We compare its clearing dynamics with CUBIC (clear, unobstructed brain imaging cocktails), a well stablished and widely used clearing method. Confocal and SPIM microscopy are used to acquire 2D and 3D images of fluorescently labelled-transparent samples. We conclude that TDE holds potential to revolutionize the study of cardiovascular pathophysiology