ASI Imaging dual selective plane illumination microscopy for cleared tissue (ct-dSPIM)

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The ASI Imaging dual selective plane illumination microscopy for cleared tissue (ct-dSPIM) allows for dual views (d) of large samples such as cleared tissue (ct).

The ct-dSPIM can be used to image various cleared tissue samples from whole mouse brains as well as slices of cleared tissue. It is one of many light sheet microscope configurations possible using ASI’s modular components. ASI manufactures the optomechanical elements, including the motorised stages and 2D galvos for creating and moving the light sheet.

ASI partnered with Special Optics to develop an objective optimised for light sheet imaging of cleared tissue. Lasers and sCMOS cameras are required to complete the system. These items can also be purchased from Scitech. Two objectives are placed at right angles above a sample mounted horizontally in an open dish, each objective 45° from vertical.

A light sheet is created from one objective and imaged using the other objective. A stack of images is collected by moving the light sheet through the sample; in the case of ct-dSPIM, the sample is normally moved through a stationary light sheet using the XY stage. For some applications, the 3D information from a single view or stack is sufficient. For dual-view systems, the role of the two objectives is reversed to collect another stack from a perpendicular direction. The two datasets can be computationally merged to yield a 3D dataset with isotropic resolution; the usual problem of poor axial resolution is overcome by information from the other view.

Other features include: image acquisition >10^8 voxels/s; sub-micron resolution in XYZ (sample-permitting); sample mounting in open dish; image >5 mm deep into flat samples or up to 12 mm radius sphere; media RI range from 1.33 to 1.56, aqueous or organic media; modular and flexible set-up.