Editors' ChoiceImaging

Seeing is believing: Tissue clearing makes see-through rodents

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Science Translational Medicine  31 Aug 2016:
Vol. 8, Issue 354, pp. 354ec138
DOI: 10.1126/scitranslmed.aah6000

Tissue clearing technologies have advanced dramatically in the past several years, enabling researchers to see deep within tissues. These tools have been particularly powerful in neuroscience, enabling tracking and tracing of neurons to map neuronal connectivity. Clearing applications for peripheral nerves, skin, and other organs have provided researchers the ability to carry out subcellular level imaging at a few hundreds of nanometers resolution throughout cubic centimeters of tissue to map cells, structures, and cellular processes.

Now, Pan et al. have pushed tissue clearing even further with a technique called ultimate DISCO (uDISCO) that clears and shrinks entire organisms for microscale imaging across even greater size scales. The same team had previously developed a method called 3DISCO, which enabled tissue clearing but resulted in a rapid loss of endogenous fluorescence. Interestingly, however, they noticed that the method caused shrinkage of tissues, which is highly advantageous for microscopy applications in which scanning over large volumes can be time consuming. The team performed a screen for organic compounds and discovered that including diphenyl ether in a mixture of solvents and antioxidants could preserve the fluorescence of green fluorescent protein (GFP) many weeks longer and at the same time induce a 65% shrinkage in tissue.

The authors demonstrated their technique by clearing entire bodies of GFP transgenic mice, enabling the cellular-level visualization of structures, including the spinal cord, optic nerve, and sciatic nerve in situ. Rats perfused with dextran-conjugated fluorophores were cleared to reveal the animals’ entire vascular networks. Importantly, the authors also verified that the uDISCO technique is compatible with both virus labeling and immunostaining, which means that the current arsenal of biological labeling tools can likely be applied alongside this tissue clearing method.

C. Pan et al., Shrinkage-mediated imaging of entire organs and organisms using uDISCO. Nat. Methods 10.1038/nmeth.3964 (2016). [Abstract]

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