Light Sheet Microscopy

This chapter focuses on the basics of the light sheet microscopy principle and construction, followed by an overview of its various implementations. It discusses some of the novel data handling solutions that light sheet microscopy gave rise to, as well as future prospects in this area. Two different ways of generating a light sheet are commonly used: in selective plane illumination microscopy (SPIM), the laser beam is expanded and focused using a cylindrical lens to form the sheet of light. Alternatively, a "virtual" light sheet can be generated by the digitally scanned light sheet microscopy (DSLM) technique, which uses a scanning mirror to rapidly sweep a beam across the field of view (FOV). The chapter discusses how to acquire 2D images of a sample using light sheet microscopy. However, most biological applications require 3D imaging. Given its excellent optical sectioning capability, speed, and low phototoxicity, light sheet microscopy is ideally suited for fast 3D imaging.