Cleared Tissue LightSheet (CTLS) is a large field light-sheet microscope designed to image whole organs at high speed. CTLS creates a focused sheet with a narrow waist for better optical sectioning, then uses a spatial light modulator (SLM) to rapidly shift the waist of the sheet along the axis of propagation.
A dual excitation setup allows imaging from the right and left sides of the specimen for optimal light-sheet projection throughout. Piezoelectric stages move the specimen in x, y, and z with sub-micron resolution. The result is clear: a 1 cm3 volume can be imaged at up to 1µm x 1µm x 3µm (XYZ) resolution, and a cleared mouse brain can be imaged in as little as 1.5 hours.
The creation of a lattice light sheet is more complex than the creation of light sheets by typical selective plane illumination microscopy (SPIM) methods. Conventional SPIM methods use an apertured Gaussian beam to form a sheet that is too thick to allow for excitation of isolated sub-cellular events.
Lattice LightSheet employs a sophisticated multi-step process to create a light sheet consisting of a parallel linear array of coherently interfering Bessel beams. The resulting light sheet is exceptionally thin and flat with unequaled optical sectioning and essentially no out-of-focus light.
Selective plane illumination (SPIM) uses a thin sheet of light to illuminate only the plane of interest, reducing phototoxicity by drastically cutting total light dose and allowing for prolonged specimen imaging over a wide range of sample sizes.
Dual inverted selective plane illumination (diSPIM) employs two orthogonal objectives positioned at 45° above the specimen plane. By alternating between the objectives for imaging and excitation, diSPIM captures two volumes that may be fused and deconvolved to achieve isotropic resolution.
Versatile mounting to a traditional inverted microscope allows for familiar sample preparation and multimodal combinations with spinning disk confocal, TIRF and photomanipulation.
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