Lattice Light Sheet Microscope

We are very excited to have the lattice light sheet microscope, designed by the lab of Eric Betzig at Janelia Research campus. Light sheet microscopy is a rapidly emerging microscopy technique that provides increased imaging speed and minimal photobleaching and photo-toxicity by using a second objective lens to restrict illumination to the focal plane of the detection objective. The lattice light-sheet microscope uses a linear array of coherent Bessel-Gauss beams to create an extremely thin light sheet that is tightly confined in the axial (Z) direction. The result is unprecedented spatiotemporal resolution when imaging live samples in three-dimensions over time, while minimizing photo-toxicity and photobleaching.

If you would like to use the lattice, please submit a request for training and we will schedule a meeting to discuss your project!

INFO FOR NEW USERS

FAQS FOR LATTICE USERS

Is lattice light sheet right for your project?

No one microscope is best for all applications. These guidelines are intended to help you decide whether the lattice light sheet might be right for your biological application.

Reduced Bleaching/Toxicity

If you are studying living cells and you struggle with photobleaching or phototoxicity when using confocal or widefield microscopes, consider LLSM. The LLSM only illuminates the focal plane of the detection objective. This reduces the photobleaching of fluorophores outside of the focal plane, and limits the dose of light to the sample.

3D Imaging

The lattice light sheet excels at rapidly acquiring full three-dimensional image stacks. By illuminating only the focal plane, the light sheet creates an “optical sectioning” effect, similar to what can be achieved with a confocal microscope but with drastically reduced photobleaching.

High Temporal Resolution

The lattice can acquire a full 3D image stack in less than a second (depending on the sample). This fact, combined with its sensitivity and light efficiency, makes the LLSM particularly well suited to the collection of three-dimensional datasets with extremely high temporal resolution.

additional considerations and limitations…

Resolution

While the LLSM is one of the highest resolution light-sheet microscopes designed, there are still limitations. Strictly speaking, one can achieve higher resolution with an oil-immersion objective on a confocal microscope. And while the LLSM does offer a super-resolution (SIM) mode, we have other SIM microscopes that can offer slightly higher resolution (though not with light-sheet). If you’re not studying a live process, and resolution is all you need, we may have better options available. Contact us to discuss your project.

Depth

The lattice is excellent at creating high contrast, optically-sectioned images with somewhat thick specimens, but there is a limit to imaging depth due to light scattering. We are always happy to try out your samples, but know that image quality will gradually degrade at many tens to hundreds of microns deep.

Field of View

Light sheet microscopy has gained a lot of attention for gorgeous movies of embryogenesis. The lattice light sheet is optimized for high resolution, but does not offer as large of a field of view as some other techniques. The largest dimension cannot be much greater than 100µm in a single field of view.

REQUEST A CONSULTATION

Lattice Filter Spectra Tool

You can evaluate the suitability of your fluorophore with the filter configuration on our lattice with this tool.

The paper

Much about the capabilities of the lattice can be learned from the original paper. Anyone interested in imaging on the lattice is strongly encouraged to familiarize themselves with the basic principle and application as described by the Betzig lab.

Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution