CUBIC-HistoVIsion: a pipeline for three-dimensional whole-organ/body staining and imaging with single-cell resolution based on chemical properties of tissue gel
The recent development of various tissue clearing and three-dimensional (3D) imaging methods, including our CUBIC technology, enabled the comprehensive observation of whole organ/body with cellular resolution or more. Several studies tried to integrate whole-mount staining into the clearing-imaging scheme. However, due to the difficulty in efficient penetration of stains and antibodies, they have only been applied in loose embryonic tissues or with a limited number of antibodies/stains for adult rodent tissues. To logically identify the critical parameters for the efficient penetration, we began by investigating material chemistry of fixed and delipidated biological tissue. Then, we performed a surrogate assay with an artificial material similar to tissue in order to widely examine multiple parameters for efficient staining. The identified parameters were integrated as a general 3D staining protocol, with which we have confirmed ~30 chemicals and antibodies used in whole adult mouse brain staining and imaging with single-cell resolution. The developed “CUBIC-HistoVIsion” pipeline for 3D histochemistry and volumetric imaging provides opportunities for multi-channel imaging of functional and structural molecules of whole adult mouse organs as well as primate organs, thus will be widely applied to life science and medical researches in future.

Susaki et al. Cell 157: 726-739 2014.
Tainaka et al. Cell 159: 911-924, 2014.
Susaki et al. Nat Protocols 10: 1709-1727, 2015.
Susaki and Ueda. Cell Chemical Biology 23:137-157, 2016. (Review)
Tatsuki et al. Neuron 90: 70-85, 2016.
Nojima et al. Sci Rep 7: 9269, 2017.
Tainaka et al. Cell Rep 24: 2196-2210.e9, 2018.
Susaki et al. in prep.
Date: 7 March 2019, 14:00 (Thursday, 8th week, Hilary 2019)
Venue: Sherrington Building, off Parks Road OX1 3PT
Venue Details: Large Lecture Theatre
Speaker: Dr Etsuo A. Susaki (The University of Tokyo Graduate School of Medicine)
Organising department: Department of Physiology, Anatomy and Genetics (DPAG)
Organiser: Professor Ana Domingos (University of Oxford, DPAG)
Organiser contact email address:
Host: Professor Ana Domingos (University of Oxford, DPAG)
Part of: Neuroscience Theme Guest Speakers (DPAG)
Booking required?: Not required
Audience: Members of the University only
Editor: Talitha Smith