High-Resolution Imaging and Morphological Phenotyping of C. elegans through Stable Robotic Sample Rotation and Artificial Intelligence-Based 3-Dimensional Reconstruction.
Peng Pan, Pengsong Zhang, Sharanja Premachandran, Ran Peng, Shaojia Wang, Qigao Fan, Yu Sun, John A Calarco, Xinyu Liu
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引用次数: 0
Abstract
Accurate visualization and 3-dimensional (3D) morphological profiling of small model organisms can provide quantitative phenotypes benefiting genetic analysis and modeling of human diseases in tractable organisms. However, in the highly studied nematode Caenorhabditis elegans, accurate morphological phenotyping remains challenging because of notable decrease in image resolution of distant signal under high magnification and complexity in the 3D reconstruction of microscale samples with irregular shapes. Here, we develop a robust robotic system that enables the contactless, stable, and uniform rotation of C. elegans for multi-view fluorescent imaging and 3D morphological phenotyping via the precise reconstruction of 3D models. Contactless animal rotation accommodates a variety of body shapes and sizes found at different developmental stages and in mutant strains. Through controlled rotation, high-resolution fluorescent imaging of C. elegans structures is obtained by overcoming the limitations inherent in both widefield and confocal microscopy. Combining our robotic system with machine learning, we create, for the first time, precise 3D reconstructions of C. elegans at the embryonic and adult stages, enabling 3D morphological phenotyping of mutant strains in an accurate and comprehensive fashion. Intriguingly, our morphological phenotyping discovered a genetic interaction between 2 RNA binding proteins (UNC-75/CELF and MBL-1/MBNL), which are highly conserved between C. elegans and humans and implicated in neurological and muscular disorders. Our system can thus generate quantitative morphological readouts facilitating the investigation of genetic variations and disease mechanisms. More broadly, our method will also be amenable for 3D phenotypic analysis of other biological samples, like zebrafish and Drosophila larvae.
期刊介绍:
Research serves as a global platform for academic exchange, collaboration, and technological advancements. This journal welcomes high-quality research contributions from any domain, with open arms to authors from around the globe.
Comprising fundamental research in the life and physical sciences, Research also highlights significant findings and issues in engineering and applied science. The journal proudly features original research articles, reviews, perspectives, and editorials, fostering a diverse and dynamic scholarly environment.