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Abstract

The goal of this PhD project was to optimize and develop new approaches for visualizing and 3D-imaging tissues and whole organs. Essentially, these approaches were based on optimizing OTC methodologies in combination to the use of CM and LSM. The core of this research was about the Ethyl Cinnamate-based OTC: starting from a recently published protocol52, we worked in order to speed up and automate all the procedures, making it suitable for further studies. Initially we started with the clearing only of mouse kidneys and, then, the procedure was extended to other tissues (normal and pathological tissues) not previously analysed yet. Additionally, we enhanced this methodology, with its application for different scopes, as “revitalization” of old paraffin samples and 3D immunohistochemistry for visualizing finer and inner structures. Our clearing protocol was also tested, for some preliminary studies, on human tissue, with promising results. In parallel, we also tried to figure out the best conditions for the use of both CM and LSM, in order to acquire 2D and 3D images for very large samples.