Preprint: This article has not been edited or peer reviewed.

Whole organ and organism tissue clearing by uDISCO

Ali Erturk, Chenchen Pan, Ruiyao Cai, Francesca Paola Quacquarelli, Alireza Gasemigharagoz
DOI: 10.1038/protex.2016.055

Abstract

As a novel organic solvent-based tissue clearing method, the uDISCO technique maintains the feature of tissue shrinkage while overcomes the fast signal quenching and preserves the endogenous fluorescence over months. By rendering the intact organs and rodent bodies transparent while reducing their size up to 65%, uDISCO enables the light-sheet microscopy of entire body of adult mice with subcellular resolution for the first time. Additionally, uDISCO is compatible with virus tracing, antibody labeling and over-fixed human tissue staining allowing the usage in various biomedical applications. Here, we provide a detailed protocol of uDISCO describing the preparation of samples and solutions, procedure of passive clearing and whole-body clearing with perfusion system. The whole procedure takes from 2 days to 1 week depending on the tissue size.

Keywords
tissue, clearing, DISCO, light-sheet, CNS, stem cells, mapping, human tissue

Figures

Introduction

Reagents

Equipment

Procedure

Acknowledgements

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Preprint: This article has not been edited or peer reviewed.

Whole organ and organism tissue clearing by uDISCO

Ali Erturk, Chenchen Pan, Ruiyao Cai, Francesca Paola Quacquarelli, Alireza Gasemigharagoz

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Abstract

As a novel organic solvent-based tissue clearing method, the uDISCO technique maintains the feature of tissue shrinkage while overcomes the fast signal quenching and preserves the endogenous fluorescence over months. By rendering the intact organs and rodent bodies transparent while reducing their size up to 65%, uDISCO enables the light-sheet microscopy of entire body of adult mice with subcellular resolution for the first time. Additionally, uDISCO is compatible with virus tracing, antibody labeling and over-fixed human tissue staining allowing the usage in various biomedical applications. Here, we provide a detailed protocol of uDISCO describing the preparation of samples and solutions, procedure of passive clearing and whole-body clearing with perfusion system. The whole procedure takes from 2 days to 1 week depending on the tissue size.

Figures

Introduction

Reagents

Equipment

Procedure

Acknowledgements

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