RT Journal Article
SR Electronic
T1 Optimization of CLARITY for Clearing Whole-Brain and Other Intact Organs
JF eneuro
JO eneuro
FD Society for Neuroscience
SP ENEURO.0022-15.2015
DO 10.1523/ENEURO.0022-15.2015
VO 2
IS 3
A1 Jonathan R. Epp
A1 Yosuke Niibori
A1 Hwa-Lin (Liz) Hsiang
A1 Valentina Mercaldo
A1 Karl Deisseroth
A1 Sheena A. Josselyn
A1 Paul W. Frankland
YR 2015
UL http://www.eneuro.org/content/2/3/ENEURO.0022-15.2015.abstract
AB The development, refinement, and use of techniques that allow high-throughput imaging of whole brains with cellular resolution will help us understand the complex functions of the brain. Such techniques are crucial for the analysis of complete neuronal morphology—anatomical and functional—connectivity, and repeated molecular phenotyping. CLARITY is a recently introduced technique that produces structurally intact, yet optically transparent tissue, which may be labeled and imaged without sectioning. However, the utility of this technique depends on several procedural variables during the process in which the light-scattering lipids in a tissue are replaced by a transparent hydrogel matrix. Here, we systematically varied a number of factors (including temperature, hydrogel composition, and polymerization conditions) to provide an optimized, highly replicable CLARITY procedure for clearing mouse brains. We found that for these preparations optimal tissue clearing requires electrophoresis (and cannot be achieved with passive clearing alone) for 5 d with a combination of 37 and 55°C temperature. Although this protocol is optimized for brains, we also show that it can be used to clear and analyze a variety of organs. Brain or other tissue prepared using this protocol is suitable for high-throughput imaging with confocal or single-plane illumination microscopy.