Monash

Discovery

CLARITY, the performance, with CUBIC variation

30.03.2016

In a nutshell: This video journal article shows how researchers use a modified CLARITY technique to see right through mice spinal cords.

View Paper Abstract
CLARITY, the performance, with CUBIC variation

The big picture:

Science can sometimes seem a mysterious art, especially with tricky techniques. One way to share tacit knowledge that is needed to replicate a complex method, but so difficult to capture in words, is to show the method rather than describe it. That’s the premise behind the Journal of Visualized Experiments (JoVE).

JOVE AL

Andy (Huazheng) Liang, a post-doc in the lab of CIBF chief investigator George Paxinos of Neuroscience Research Australia and the University of New South Wales in Sydney, “performs” the CLARITY technique in a JoVE video paper.

CLARITY — it stands for Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining / in situ-hybridization-compatible Tissue hYdrogel— renders the mouse spinal cord into a transparent, stainable structure ready for 3-dimensional analysis in exquisite detail.

First, the spinal cord is infused with a hydrogel solution, which acts as transparent scaffold, anchoring the proteins and nucleic acids in place. Then a series of detergent washes remove fats from the tissue, making it transparent enough to read 8pt font through. Next, fluorescent antibodies are used to tag molecules of interest, such as serotonin. Finally, a “light sheet” fluorescent microscope, which can quickly scan three-dimensional pieces of tissue, is used to visualize the molecules within and around neurons and nerve fibres.

The whole process can take from a week to a couple of months, depending on the protocol used. To speed up the fat-clearing process, Liang has incorporated a variation into the technique called CUBIC.

JoVE’s papers are peer-reviewed, and show techniques within the context of an experiment — in this case, Liang is studying connections between the mouse hindbrain and spinal cord.

Video, says JoVE editorial director Avital Braiman, improves the reproducibility of results—long a roadblock to scientific progress. “Science can’t advance without reproducible results,” she says.


Reference:
Liang, H., Schofield, E., & Paxinos, G. (2016). Imaging Serotonergic Fibers in the Mouse Spinal Cord Using the CLARITY/CUBIC Technique. JoVE (Journal of Visualized Experiments), (108), e53673-e53673.


Republish this article:

We believe in sharing knowledge. We use a Creative Commons Attribution 4.0 International License, which allows unrestricted use of this content, subject only to appropriate attribution. So please use this article as is, or edit it to fit your purposes. Referrals, mentions and links are appreciated.

CIBF