CLARITYA technique for structural and molecular interrogation of intact biological systems

Based on the work of Kwanghun Chung and Karl DeisserothPublished on Nature Methods 2013

Studying intact systems with molecular resolution and global scope remains an unmet

goal in biology

CONFOCALTWO-PHOTONmax penetration

700 um

Packed LIPID bilayers

Diffusion Barrier

Light ScatteringPOOR IMAGING!

NO AB PENETRATION!

Liposome

Micelle

Bilayer sheet

GOALTransform intact tissue into an optically

transparent and macromolecule-permeable construct while preserving molecular information

and structure.

CLARITY the method

Chung & Deisseroth (2013)

CLARITY the method

Chung & Deisseroth (2013)

CLARITY works!

Chung et al. (2013)

EXAMPLE whole mice brain

Chung et al. (2013)

EXAMPLE whole mice brain

Chung & Deisseroth (2013)

EXAMPLE zebra fish

Chung & Deisseroth (2013)

EXAMPLE human brain

Chung & Deisseroth (2013)

EXAMPLE multi-labeling

Chung & Deisseroth (2013)

Tomer et al. (2014)

EXAMPLE mice brain LSM

Tomer et al. (2014)

EXAMPLE mice brain LSM

EXAMPLE 3D rendering

Confocal Light Sheet

Tomer et al. (2014)Chung & Deisseroth (2013)

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

Poguzhelskaya et al. (2014)

CLARITY opportunities

➲ Application of CLARITY to other tissues

➲ Combination with other techniques

➲ Functional mesh design

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