University of Minnesota
University Imaging Center
http://uic.umn.edu/
612.624.7938
University Imaging Centers - Where the University sees the future

Tissue Clearing

The University Imaging Centers integrates the most recent clearing tissue and imaging technologies in order to image and analyze intact tissues and organs in three-dimensions.

Tissue clearing has been developed in multiple laboratories to make tissues optically transparent to optimize whole-mount imaging. It also enables internal structure preservation and permeability to macromolecules like stains and antibodies. The UIC will have multiple clearing and labeling approaches with X-Clarity being the most widely useful. With the X-Clarity method tissue structure is immobilized in a hydrogel to study fluorescent labels in biological systems in 3D. This approach brings a new dimension to histology services. This step can process samples of various sizes (samller the better with < 8 cubic mm optimal, although larger may be possible and is limited by imaging) and we have the sample holders for for 6 Mouse Brain sized samples, 2 Rat Brain sized samples, tissue slices (<1.5mm each) and 48 tissue samples (<3 cubic mm each) with many other options.

As for imaging, the UIC is equipped with a 3i Cleared Tissue Light-Sheet, Caliber RSG4 ribbon scanning, confocal microscopy, multiphoton microscopy and light sheet fluorescence microscopy. We can work with you to determine which system and which probes are right for you.

This approach enables the conservation and imaging of proteins and nucleic acids throughout intact tissue. It offers the opportunity to show the relationship between structure and function of a molecule of interest in an intact organ.

The UIC will continue to expand tissue clearing and 3D imaging service to multiple fields of research in samples ranging from plants to animals.

Fixation and sample prep for your samples for submission

Mouse brain

perfusion d1X PBS and then 4% PFA, followed by brain extraction and immersion in 4% PFA for 24h at 4C. Brains should then be washed with 1X PBS and stored in PBS at 4C. After this you may drop off the samples for clearing.

 

Questions about clearing?

Please contact Mark Sanders (msanders@umn.edu), and Mary Brown (mebrown@umn.edu).

 

Click here to submit a 

Tissue Clearing Request

 

X-CLARITY Tissue Clearing System

The X-CLARITY is a collection of systems and ready-to-use reagents to standardize, simplify, and accelerate each step of the tissue clearing process. X-CLARITY is based on CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining / in situ-hybridization-compatible Tissue hyYdrogel) method. With CLARITY, preserved tissues are embedded in a hydrogel matrix and lipis are actively extracted through electrophresis to create a stable and optically transparent tissue-hydrogel hybrid that is chemically accessible for multiple rounds of antibody labeling and imaging.

 
 
     

     

X-CLARITY Workflow