CUBIC-HistoVIsion: a pipeline for three-dimensional whole-organ/body staining and imaging with single-cell resolution based on chemical properties of tissue gel
Mar. 18, 2019 16:00 - 17:00

Seminar

Kobe

DB Bldg. SeminarRoom A7F

Etsuo Susaki

The University of Tokyo, Department of Systems Pharmacology, Graduate school of medicine

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Recent development of various tissue clearing and three-dimensional(3D) methods 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 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 chemical 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 histology 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.

Kazunari Miyamichi