Developers
Bin Yang, Jennifer B. Treweek, Long Cai, Viviana Gradinaru, etc.
Description of the technology
The proposed technology enables to visualize the cellular architecture of entire organs using techniques for making tissues transparent, allowing to see organs and inner structures of animals at depths of millimeters to centimeters by using microscopes.
For «optical clearing» of animal tissues, the special technique is applied, named CLARITY, which in original version involves the stabilization of specimens with an acrylamide hydrogel and then the depletion of them with lipids. In the frameworks of this technology, the modified version of CLARITY is used. In process of this modified version, hydrogel monomers and
The technology includes PACT, a protocol for passive tissue clearing and immunostaining of intact organs; RIMS, a refractive index matching media for imaging thick tissue; and PARS, a method for
The demonstration of applicability of the technology was carried out by the clearing of a variety of mouse organs, including brain, heart, lung, intestine and kidney as well as the body of a whole mouse. It was shown thereby that PACT, RIMS, and PARS are compatible with
Practical application
The technology allows the observing of cells and molecules within the intact architecture of the tissues and organs, which is crucial for understanding of the structure and function of complex biological systems. It is applicable to identify the
Besides, the technology enables systematic identification of cell types throughout the volume monitored during previous live imaging. This identification is carried out by staining of key transcription factors and other genes of interest or by
Laboratories
- Division of Biology and Biological Engineering, California Institute of Technology, Pasadena (USA)
- Division of Dermatology, Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles (USA)
- Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena (USA)
Links
http://www.nature.com/nbt/journal/v32/n11/full/nbt.3061.htmlhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4153367/
Publications
- Höckendorf, B., Lavis, L.D., Keller, P.J. «Making biology transparent» 32.11 Nature Biotechnology, (2014): 1104–1105.
- Yang, B. et al. «
Single-cell phenotyping within transparent intact tissue throughwhole-body clearing." 158.4 Cell. (2014): 945−958. - Chung, K. et al. «Structural and molecular interrogation of intact biological systems." 497 Nature, (2013): 332−337.
- Gradinaru,
v. et al. «Molecular and cellular approaches for diversifying and extending optogenetics." 141.1 Cell. (2010): 154−165.