Speaker
Description
We have established long-term light sheet imaging of human brain organoid development for the spatiotemporal profiling of developmental morphodynamics underlying human brain patterning and morphogenesis. Here, we present and analyze a novel dual-channel, multi-mosaic and multi-protein labeling strategy combined with a demultiplexing approach. To achieve this, we have developed Light-Insight, a novel computational workflow, leveraging morphological measurements and machine learning to demultiplex the different labelled fluorescent cell lines. We simultaneously profile and track Actin, Tubulin, plasma membrane, Histone, and LAMB1 dynamics. We apply Light-Insight to monitor week-long human brain organoid development and unravel the impact of extrinsically provided extracellular matrix. Our workflow enables quantifying cell morphologies, alignment changes and tissue scale dynamics during tissue state transitions including neuroepithelial induction, lumenization, and brain regionalization. We show that an extrinsically provided matrix enhances lumen expansion and cell elongation, while the intrinsic matrix or an encapsulation of the neural organoids in agarose have altered cell and tissue morphologies. Overall, Light-Insight and our imaging approach quantify tissue state transitions and can be applied to any healthy or perturbed developmental or organoid system.
| Authors | Gilles Gut*, Akanksha Jain*, Ryoko Okamoto, J. Gray Camp, Barbara Treutlein |
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| Keywords | Light sheet imaging, spatiotemporal profiling, morphodynamics, demultiplexing, tissue dynamics, cell morphology, human brain organoids |
