Tissue Clearing: An Overview of Methods, Considerations, and Applications

Abstract Since the advent of confocal microscopy, advances in tissue clearing have allowed researchers to push the limits of imaging depth and dive deeper into the precise 3D structures of biological specimens. As clearing methods are discovered, refined, and retired, it behooves the prudent microscopist to consider the pros and cons of various tissue clearing methods and systematically evaluate whether tissue clearing is beneficial to answer their research questions. Herein, I will briefly discuss the evolution of tissue clearing, pros and cons of more popular methods and techniques related to fluorescence microscopy, and key considerations for selecting a method most likely to yield experimental success.