Applied Scientific Instrumentation releases dual selective plane illumination microscopy

(Photo Courtesy of Applied Scientific Instrumentation)

The ct-dSPIM is a flexible and easy-to-use selective plane illumination microscopy (SPIM) configuration from Applied Scientific Instrumentation (ASI), made from the company’s flexible and modular microscope components, allowing dual-views of large samples of cleared tissue.

When used with ASI and Special Optics’s new objective optimized for light sheet imaging of cleared tissue, imaging depth can exceed 5mm 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.

As a dual-view system, the roles of the objectives can then be reversed to collect another stack from a different perspective; computationally merging the two stacks yields a 3D dataset with ~2x improvement in axial resolution.

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