Light sheet fluorescence microscopy imaging of cleared tissues

The unique optical design used in light sheet fluorescence microscopy (LSFM) (aka SPIM ) allows for imaging of very thick specimens at much greater speeds than other fluorescence microscopy methods. Recent advances in tissue clearing methods have made it possible to perform such imaging on samples that are naturally opaque. Tissues such as whole rodent brains can be rendered clear, labelled with fluorescent markers or antibodies, and imaged intact.  As opposed to sectioning, all relational and positional information is retained, making it possible to see the three-dimensional relationships of cells and structures within the tissues.  Imaging a cleared mouse hemi-brain can be performed in about an hour, many times faster than confocal microscopy.

Thanks to the Office of the VPR, we are now able to offer optimized cleared sample imaging for the Zeiss Lightsheet Z.1. The optics accommodate clearing methods with a refractive index of approximately 1.45, including CLARITY, LUMOS, Scale-S, CUBIC2 and several other techniques. The Lightsheet Z.1 is an ideal imaging option for many projects using cleared specimens, including those prepared using the X-Clarity active clearing system available in the Light Microscopy Core. The Z.1 sample chamber can accept and image tissues as large as a mouse brain (see Zeiss movie of EGFP-labelled brain and images from this facility of rodent brain vasculature). Using the 5x objective with 2.5 fold zoom, an effective magnification of up to 125x can be achieved. Multiple individual Z-stacks (tiles) may be collected and stitched together to create a single 3D rendering of the entire tissue.

For more information about LSFM, cleared tissue imaging, or to discuss a potential project, please contact us.

(Mouse brain vasculature image courtesy of David Braun.)

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