preparation-and-live-imaging-of-regenerating-hydra-spheroids

Preparation and Live Imaging of Regenerating <em>Hydra</em> Spheroids

Hydra is renowned for its exceptional regenerative capacity. Remarkably, small regenerating fragments of Hydra tissue, called Hydra spheroids, can ...

Friedrich Miescher Institute for Biomedical Research

University of Basel

Multicellular systems grow over the course of weeks from single cells to tissues or even full organisms, making live imaging challenging. To bridge spatiotemporal scales, we present an open-top dual-view and dual-illumination light-sheet microscope dedicated to live imaging of large specimens at single-cell resolution. The configuration of objectives together with a customizable multiwell mounting system combines dual view with high-throughput multiposition imaging. We use this microscope to image a wide variety of samples and highlight its capabilities to gain quantitative single-cell information in large specimens such as mature intestinal organoids and gastruloids.

Open-top multisample dual-view light-sheet microscope for live imaging of large multicellular systems.

The spontaneous emergence of tissue patterns is often attributed to biochemical reaction-diffusion systems. In  tissue regeneration, the formation of a Wnt signaling center exemplifies this process. However, a strictly biochemical mechanism for self-organization in  remains elusive. In this study, we investigated mechanical stimuli and identified a positive feedback loop between Wnt signaling and tissue stretching. We developed a mathematical model of mechanochemical pattern formation in a closed elastic shell, representing regenerating  epithelial spheroids. Our model explains how mechanical forces drive axis formation and predict the organizer's location under various perturbations. Validation by partially confining regenerating tissues showed that the organizer forms in regions with the greatest stretching potential. This work highlights a versatile mechanochemical mechanism for luminal epithelium patterning, which is relevant across various biological systems.

Article	Year
Open-top multisample dual-view light-sheet microscope for live imaging of large multicellular systems. Nature methods\| PubMed ID: 38509326	2024
Mechanochemical patterning localizes the organizer of a luminal epithelium. Science advances\| PubMed ID: 40561030	2025

Sera L. Weevers

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