Imaging three-dimensional brain organoid architecture from meso- to nanoscale across development

Author:

Rodriguez-Gatica Juan Eduardo1,Iefremova Vira2,Sokhranyaeva Liubov3,Yeung Si Wah Christina Au2,Breitkreuz Yannik4,Brüstle Oliver24ORCID,Schwarz Martin Karl34ORCID,Kubitscheck Ulrich1ORCID

Affiliation:

1. Institute of Physical and Theoretical Chemistry, University of Bonn 1 , Wegelerstr. 12, 53115 Bonn , Germany

2. Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital Bonn 2 , Venusberg-Campus 1, 53127 Bonn , Germany

3. Institute of Experimental Epileptology and Cognition Research (IEECR), University of Bonn Medical School 3 , Sigmund-Freud-Str. 25, 53127 Bonn , Germany

4. LIFE & BRAIN GmbH 4 , Cellomics Unit, Venusberg-Campus 1, D-53127 Bonn , Germany

Abstract

ABSTRACT Organoids are stem cell-derived three-dimensional cultures offering a new avenue to model human development and disease. Brain organoids allow the study of various aspects of human brain development in the finest details in vitro in a tissue-like context. However, spatial relationships of subcellular structures, such as synaptic contacts between distant neurons, are hardly accessible by conventional light microscopy. This limitation can be overcome by systems that quickly image the entire organoid in three dimensions and in super-resolution. To that end we have developed a system combining tissue expansion and light-sheet fluorescence microscopy for imaging and quantifying diverse spatial parameters during organoid development. This technique enables zooming from a mesoscopic perspective into super-resolution within a single imaging session, thus revealing cellular and subcellular structural details in three spatial dimensions, including unequivocal delineation of mitotic cleavage planes as well as the alignment of pre- and postsynaptic proteins. We expect light-sheet fluorescence expansion microscopy to facilitate qualitative and quantitative assessment of organoids in developmental and disease-related studies.

Funder

Deutsche Forschungsgemeinschaft

Deutscher Akademischer Austauschdienst

Agencia Nacional de Investigación y Desarrollo

European Commission

Publisher

The Company of Biologists

Subject

Developmental Biology,Molecular Biology

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