ABSTRACT
In recent years, a variety of optical clearing methods have emerged as powerful tools for three-dimensional imaging of biological tissues. Organic solvent–based clearing methods, such as 3DISCO, have the advantage of high clearing performance and substantial size reduction for panoptic imaging of large samples such as whole organs and even whole bodies. However, the loss of endogenous fluorescence remains a major concern for solvent-based clearing methods. To overcome this issue, researchers have approached the problem from different perspectives. This chapter describes some related work to improve endogenous fluorescence using different strategies, involving the introduction of a resin formulation during archiving of cleared samples, screening the fluorescence-friendly compounds, and adjusting the conditions in the clearing and storage process. These advanced methods have provided useful and valuable alternatives for the long-term preservation of endogenous fluorescence, facilitating the three-dimensional mapping of whole organs with the potential to be extensively used in biomedical studies.
