May 15, 2018 (Vol. 38, No. 10)
The ct-dSPIM is a flexible and easy-to-use Selective Plane Illumination Microscopy (SPIM) configuration made from flexible and modular microscope components, allowing dual-views of large samples of cleared tissue. When used with ASI’s and Special Optics’s new objective optimized for light-sheet imaging of cleared tissue, imaging depth can exceed 5 mm into flat samples. The objective accommodates media RI from 1.33 to 1.56 (aqueous or organic media). The ct-dSPIM has successfully imaged various cleared tissue samples from microtome-cut slices to whole mouse brains. Unlike other light sheet techniques, this SPIM concept uses two objectives, oriented 45° from vertical, at right angles to one another above a horizontally mounted sample in an open dish. A light sheet is created from one objective and imaged through the other. A stack of images can be collected by moving the light sheet through the sample. In the case of the ct-dSPIM, the sample is normally moved through a stationary light sheet using an XY stage.