Current imaging approaches limit the ability to perform multiscale characterization of 3D organotypic cultures (organoids) in large numbers. Here, we present an automated multiscale 3D imaging platform synergizing high-density organoid cultures with 3D live single objective light-sheet imaging. It is composed of disposable microfabricated organoid culture chips embedding optical components and a custom laser beam steering unit coupled to a commercial inverted microscope. It streamlines organoid culture and high content 3D imaging on a single user-friendly instrument with minimal manipulations and unprecedented throughput of 300 organoids per hour in 3D. Collecting large number of 3D stacks allowed training deep learning-based algorithms to quantify the organoids morphogenetic organizations at multi-scales, ranging from the sub-cellular scale to the whole organoid level.We validated the versatility and robustness of our approach on intestine, hepatic, neuroectoderm organoids and oncospheres.
A controlled ''green synthesis'' approach to synthesize silver nanoparticles by Allium cepa and Musa acuminata plant extract has been reported. The effect of different process parameters, such as pH, temperature and time, on synthesis of Ag nanoparticles from plant extracts has been highlighted. The work reports an easy approach to control the kinetics of interaction of metal ions with reducing agents, stabilized by ammonia to achieve sub-10 nm particles with narrow size distribution. The nanoparticles have been characterized by UV-Visible spectra and TEM analysis. Excellent antimicrobial activity at extremely low concentration of the nanoparticles was observed against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Fusarium oxysporum which may allow their exploitation as a new generation nanoproduct in biomedical and agricultural applications.
The generation of structurally standardized human pluripotent stem cell (hPSC)‐derived neural embryonic tissues has the potential to model genetic and environmental mediators of early neurodevelopmental defects. Current neural patterning systems have so far focused on directing cell fate specification spatio‐temporally but not morphogenetic processes. Here, the formation of a structurally reproducible and highly‐organized neuroepithelium (NE) tissue is directed from hPSCs, which recapitulates morphogenetic cellular processes relevant to early neurulation. These include having a continuous, polarized epithelium and a distinct invagination‐like folding, where primitive ectodermal cells undergo E‐to‐N‐cadherin switching and apical constriction as they acquire a NE fate. This is accomplished by spatio‐temporal patterning of the mesoendoderm, which guides the development and self‐organization of the adjacent primitive ectoderm into the NE. It is uncovered that TGFβ signaling emanating from endodermal cells support tissue folding of the prospective NE. Evaluation of NE tissue structural dysmorphia, which is uniquely achievable in the model, enables the detection of apical constriction and cell adhesion dysfunctions in patient‐derived hPSCs as well as differentiating between different classes of neural tube defect‐inducing drugs.
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