Passive and Whole Body CLARITY for Single Cell Phenotyping

Speakers: Dr Benjamin E. Deverman, PhD (Senior Research Scientist, California Institute of Technology) and Arvonn Tully (Advanced Application Scientist, Bitplane)

Understanding the structure-function relationships at cellular, circuit, and organ-wide scale requires 3D anatomical and phenotypical maps, currently unavailable for many organs across species. At the root of this knowledge gap is the absence of a method that enables whole-organ imaging. Recently we presented techniques for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically transparent, thereby exposing their cellular structure with intact connectivity (Yang et al 2014).

Therein we described PACT (passive clarity technique), a protocol for passive tissue clearing and immunostaining of intact organs; RIMS (refractive index matching solution), a mounting media for imaging thick tissue; and PARS (perfusion-assisted agent release in situ), a method for whole-body clearing and immunolabeling. We showed that in rodents PACT, RIMS, and PARS are compatible with endogenous-fluorescence, immunohistochemistry, RNA single-molecule FISH, long-term storage, and microscopy with cellular and subcellular resolution.

These methods are applicable for high-resolution, high-content mapping and phenotyping of normal and pathological elements within intact organs and bodies. In this webinar we will provide an overview of these techniques.

Key learning objectives:

• Basics and application of a novel whole body tissue clearing technique – PACT (passive clarity technique)
• Imaris Software as a tool for visualisation and morphology analysis of the structures