Based on the work of Kwanghun Chung and Karl Deisseroth, CLARITY technique allows to clarify thick tissues to make them suitable for diverse microscopy techniques.

1. CLARITY
A technique for structural and molecular
interrogation of intact biological systems
Based on the work of Kwanghun Chung and Karl Deisseroth
Published on Nature Methods 2013

2. Studying intact systems with molecular
resolution and global scope remains an unmet
goal in biology
CONFOCALTWO-PHOTON
max penetration
700 um

3. Packed LIPID bilayers
Diffusion Barrier
Light Scattering
POOR IMAGING!
NO AB PENETRATION!
Liposome
Micelle
Bilayer sheet

4. GOALTransform intact tissue into an optically
transparent and macromolecule-permeable
construct while preserving molecular information
and structure.

5. CLARITY the method
Chung & Deisseroth (2013)

6. CLARITY the method
Chung & Deisseroth (2013)

7. CLARITY works!
Chung et al. (2013)

8. EXAMPLE whole mice brain
Chung et al. (2013)

9. EXAMPLE whole mice brain
Chung & Deisseroth (2013)

10. EXAMPLE zebra fish
Chung & Deisseroth (2013)

11. EXAMPLE human brain
Chung & Deisseroth (2013)

12. EXAMPLE multi-labeling
Chung & Deisseroth (2013)

13. Tomer et al. (2014)

14. EXAMPLE mice brain LSM
Tomer et al. (2014)

15. EXAMPLE mice brain LSM

16. EXAMPLE 3D rendering
Confocal Light Sheet
Tomer et al. (2014)Chung & Deisseroth (2013)

17. CLARITY not so clear? CHALLENGES
➲ Can’t be applied to live tissue
➲ Time consuming protocol
➲ High technical skills required
➲ Long working distance objectives
➲ High amount of data generated

18. Poguzhelskaya et al. (2014)

19. CLARITY opportunities
➲ Application of CLARITY to other tissues
➲ Combination with other techniques
➲ Functional mesh design

20. Chung, K., & Deisseroth, K. (2013). CLARITY for mapping the nervous system. Nature
methods, 10(6), 508-513.
Chung, K., Wallace, J., Kim, S. Y., Kalyanasundaram, S., Andalman, A. S., Davidson, T. J.,
Mirzabekov, J. J., Zalocusky, K. A., Mattis, J., Denisin, A. K., Pak, S., Bernstein, H.,
Ramakrishnan, C., Grosenick, L., Gradinaru, V. & Deisseroth, K. (2013). Structural and
molecular interrogation of intact biological systems. Nature, 497(7449), 332-337.
Poguzhelskaya, E., Artamonov, D., Bolshakova, A., Vlasova, O., & Bezprozvanny, I. (2014).
Simplified method to perform CLARITY imaging. Molecular neurodegeneration, 9(1), 19.
Tomer, R., Ye, L., Hsueh, B., & Deisseroth, K. (2014). Advanced CLARITY for rapid and
high-resolution imaging of intact tissues. nature protocols, 9(7), 1682-1697.
Ke, M. T., Fujimoto, S., & Imai, T. (2013). SeeDB: a simple and morphology-preserving
optical clearing agent for neuronal circuit reconstruction. Nature neuroscience, 16(8),
1154-1161.
CLARITY Forum. Available at: http://forum.claritytechniques.org. Last accessed 26 Jan
2015.
CLARITY references