Light Sheet Fluorescence Microscopy Combined with Optical Clearing Methods as a Novel Imaging Tool in Biomedical Research

Abstract

In the past decade, imaging has advanced to become a crucial tool in fundamental and biomedical research and it has become increasingly important to be able to image whole organs with single cell resolution. Light sheet fluorescence microscopy, also called selective plane illumination microscopy or ultramicroscopy, provides a high resolution in transparent and intact whole organs. By the application of a thin light sheet, only a defined slice of the specimen is illuminated and the fluorescence signal is detected by an objective perpendicular to the specimen. By moving the specimen vertically through the laser, a z-stack is acquired which corresponds to an optical sectioning without physical disruption of the specimen. The data can further be reconstructed to a three-dimensional volume and analysed in its entire complexity in micrometre resolution. This article reviews the prerequisites for successful light sheet fluorescence microscopy, in terms of tissue preparation and optical clearing, and highlights recent advances and applications in the context of basic and biomedical research, with special focus on the central nervous system of rodents.