Using Light Sheet Imaging for Fluorescence Microscopy

Light sheet fluorescence microscopy (LSFM) is a powerful tool in the life science researcher’s toolkit. While the old histopathology workflow of physical sectioning and slide mounting still remains relevant, LSFM continues to shift the paradigm of imaging whole tissues, organs and organisms (via optical sectioning of cleared specimens). With powerful tools like LSFM and the Visikol® HISTO™ clearing reagents, researchers can capture high content images without damaging or distorting tissue morphology.

Light sheet microscopes work by illuminating samples not with a one-dimensional laser point, but with a two dimensional sheet of light. When two of the three spatial dimensions are illuminated at once, the image capturing speeds can be orders of magnitude faster than single point scanning platforms like confocal. Less illuminations means less risk of photobleaching precious samples. The excitation laser sheet is emitted perpendicularly to the objective to allow for the collection of all emission within this illumination plane.

The light sheet is great at avoiding out of focus signals, which increases sensitivity by contrast optimization. One of the greatest advantages of LSFM is the ability to 3D image specimens generally considered too large for conventional fluorescence microscopes. From organoids to whole animals, the flexibility in specimen range makes this platform adaptable for many sample sizes. LSFM utilizes both air and immersion objectives for both high throughput 4x scans and high resolution 20x scans.

Light sheet fluorescence microscopy is not just for large samples. 3D cell culture models are also amenable to LSFM scanning. A spheroid for example has a small size but a great deal of complexity, with the interactions between neighboring cells often determining the relevancy of in vivo disease states. LSFM can rapidly map these interactions without sacrificing the resolution needed to glean real insight into the significant changes in structure of these models. Structural fidelity is a critical goal of many disease model-based biotechnologies, including more cutting-edge platforms 3D bio-printing. Biological structures made by 3D bio-printing must have tight tolerances at microscopic scales. The required quality control of such models make suitable for optical sectioning with LSFM.

Visikol has launched light sheet microscopy services in partnership with Bruker. We are now offering a range of services from neural network mapping to quantifying volumes of vasculature in any organ. If your research could benefit from a greater understanding of the structural conditions present in your in-vivo specimens, Visikol’s imaging and digital pathology expertise paired with Bruker’s Luxendo MuVi SPIM LSFM platforms can provide those insights and more.

Reach out today to speak with one of our knowledgeable scientists who will find the best way to apply Visikol’s leading imaging capabilities to accomplishing your research goals.

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