Light Sheet Microscopy
Oocyte labelled for tubulin and actin captured with Marianas LightSheet™.
Light Sheet Microscopy
Oocyte captured with Marianas LightSheet
The Difference is Clear
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.
Smooth muscle cells in the arteries, veins, and capillaries of mouse brain cleared with PEGASOS labeled by NG2BacDsRed.
Sample courtesy of Dr. Woo-Ping Ge, University of Texas Southwestern Medical Center.
Smooth muscle cells in the arteries, veins, and capillaries of mouse brain cleared with PEGASOS labeled by NG2BacDsRed.
Sample courtesy of Dr. Woo-Ping Ge, University of Texas Southwestern Medical Center.
Extremely Efficient Illumination
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.
HeLa cells transfected with GFP-Lifeact (488nm) to visualize actin dynamics and stained with DiL (561nm red)(Thermo) to visualize membrane dynamics.
Courtesy of James Springfield, Institute for Molecular Bioscience (IMB) at the University of Queensland
HeLa cells transfected with GFP-Lifeact (488nm) to visualize actin dynamics and stained with DiL (561nm red)(Thermo) to visualize membrane dynamics.
Courtesy of James Springfield, Institute for Molecular Bioscience (IMB) at the University of Queensland
Multimodal dual view imaging with traditional specimen preparation
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.
