CUBIC-HistoVIsion: a versatile three-dimensional whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissue

Optical – Keynote Session
12:30 – 13:05 (Sydney Time) | Tuesday 16 Feb 2020

Abstract

The recent development of various tissue clearing and three-dimensional (3D) imaging methods, including our CUBIC pipeline [1-2], allowed the comprehensive observation of the whole organ/body with cellular resolution or more. However, in the long history of histology, whole-organ/body 3D staining and imaging have been challenging due to the difficulty of adequate penetration of stains and antibodies. Even a small dye occasionally exhibits resistance to penetration, implying a complex physicochemical environment in the staining system.

In this presentation, we will introduce a versatile whole-organ/body staining and imaging protocol named CUBIC-HistoVIsion [3]. To dissect the complex physicochemical environment, we first conducted a precise characterization of biological tissue as an electrolyte gel. Then, we experimentally evaluated a broad range of 3D staining conditions by using a simplified tissue-mimicking artificial electrolyte gel. The combination of essential conditions allowed a bottom-up design of efficient 3D staining protocol which could uniformly label adult whole mouse brains, an adult marmoset hemisphere, a ~1 cm3 tissue block of adult human postmortem cerebellum, and an infant whole marmoset body with dozens of antibodies and cell-impermeant nucleic acid stains. We also demonstrate that our protocol enabled structural and functional neural circuit identification and analysis with Rabies virus tracing and whole-brain c-Fos immunostaining. The CUBIC-HistoVIsion offers advanced opportunities for organ- and organism-scale histological analysis of multicellular systems in the brain and body.

Reference:
1. Susaki et al. Cell 157: 726-739 (2014)
2. Susaki et al. Nature Protocols 10: 1709-1727 (2015)
3. Susaki et al. Nature Communications 11: 1982 (2020)

Speaker

Dr Etsuo A. Susaki

Dr Etsuo A. Susaki

Associate Professor, The University of Tokyo Graduate School of Medicine

Dr. Etsuo A. Susaki graduated from Kyushu University School of Medicine in 2002 and received his Ph.D. (Medicine) in 2007. His research focuses on the functional state of multicellular systems and their regulatory mechanisms. In particular, he developed the CUBIC framework and realized a cell-omics approach for studying multicellular systems. More recently, he has also successfully developed CUBIC-HistoVIsion, an efficient 3D staining and imaging technique for whole organs and the whole body. He is also an expert in molecular biology, biochemistry, and genetics. He is currently using his unique research background and techniques to uncover the early-stage processes of disease and aging.