Visualizing fast 3D dynamics of biological samples using light-sheet microscopy

Light sheet fluorescence microscopy (LSFM) is a convenient tool for high resolution, 3D bio-imaging as it efficiently collects the generated fluorescence while at the same time minimizes photobleaching. Being based on an intrinsic plane illumination, it allows for a fast 2D imaging. Therefore, LSFM has been put forward as an interesting candidate for fast volumetric (3D) imaging.Here I will present a LSFM microscope, combined with the use of wavefront coding (WFC) techniques, for fast volumetric imaging. WFC is used to extend the depth of field (DOF) of the collecting objective in a LSFM. This result in a system in which the light sheet can be scanned through the sample, which remains static, providing the LSFM with intrinsic 3D imaging capabilities. In addition, because of the extended DOF, the light sheet can be axially scanned at fast speeds. As only the light sheet is moved, fast 3D imaging can be achieved without the need of any sample or objective movement.