The ct-dSPIM is a flexible and easy-to-use implementation of Selective Plane Illumination Microscopy (SPIM) that allows for dual views (d) of large samples such as cleared tissue (ct).

The ct-dSPIM is one of many light sheet microscope configurations possible using ASI’s modular components. We manufacture the optomechanical elements, including the motorized stages and 2D galvos for creating and moving the light sheet. ASI partnered with Special Optics to develop an objective optimized for light sheet imaging of cleared tissue. Lasers and sCMOS cameras are required to complete the system; users can procure these themselves, use the services of various system integrators selling ASI SPIM systems, or purchase them via ASI.

The ct-dSPIM has been successfully used to image various cleared tissue samples including whole mouse brains and slices of cleared tissue.

Single-Sided System (iSPIM)

Light sheet created from one objective and imaged using the other objective.

Advantages: Fastest acquisition, least expensive.

Disadvantages: Better XY resolution than Z resolution.

Dual-Sided System (diSPIM)

Both objectives have a light sheet scanner and camera. Data is collected from both views sequentially, and the two datasets can be merged computationally to yield a 3D dataset with isotropic resolution. Can operate in single-sided mode if desired.

Advantages: XY and Z resolutions are all very good.

Disadvantages: More hardware to buy. Isotropic resolution requires double the acquisition time plus data post-processing.

ct-SPIM Example Variations

• Filter wheels on imaging paths

• Asymmetric single-sided system, e.g. using other manufacturer’s high-NA clearing objective lens

• Non-gaussian beam

• 2-photon microscopy

ct-dSPIM Features

• Image acquisition >10⁸ 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 setup