Quantitative studies of cellular morphodynamics rely on accurate cell segmentation in live cell images. However, fluorescence and phase contrast imaging hinder accurate edge localization. To address this challenge, we developed MARS-Net, a deep learning model integrating ImageNet-pretrained VGG19 encoder and U-Net decoder trained on the datasets from multiple types of microscopy images. Here, we provide the protocol for installing MARS-Net, labeling images, training MARS-Net for edge localization, evaluating the trained models' performance, and performing the quantitative profiling of cellular morphodynamics. For complete details on the use and execution of this protocol, please refer to Jang et al. (2021).
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http://dx.doi.org/10.1016/j.xpro.2022.101469 | DOI Listing |
Elife
January 2025
CNRS UMR168, Institut Curie, Université PSL, Sorbonne Université, Paris, France.
Live-cell microscopy routinely provides massive amounts of time-lapse images of complex cellular systems under various physiological or therapeutic conditions. However, this wealth of data remains difficult to interpret in terms of causal effects. Here, we describe CausalXtract, a flexible computational pipeline that discovers causal and possibly time-lagged effects from morphodynamic features and cell-cell interactions in live-cell imaging data.
View Article and Find Full Text PDFDev Cell
January 2025
Premium Research Institute for Human Metaverse Medicine (WPI-PRIMe), Graduate School of Medicine, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita-shi, Osaka 565-0871, Japan; Human Biology Research Unit, Institute of Integrated Research, Institute of Science Tokyo, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; Divisions of Gastroenterology, Hepatology & Nutrition, and Developmental Biology, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA; Department of Pediatrics, University of Cincinnati College of Medicine, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA; Center for Stem Cell and Organoid Medicine (CuSTOM), Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA. Electronic address:
Recent advancements in pluripotent stem cell and synthetic tissue technology have brought significant breakthroughs in studying early embryonic development, particularly within the first trimester of development in humans. However, during fetal stage development, investigating further biological events represents a major challenge, partly due to the evolving complexity and continued interaction across multiple organ systems. To bridge this gap, we propose an "in toto" biological framework that leverages a triad of technologies: synthetic tissues, intravital microscopy, and computer vision to capture in vivo cellular morphodynamics, conceptualized as single-cell choreography.
View Article and Find Full Text PDFSTAR Protoc
January 2025
Gladstone Institutes, San Francisco, CA, USA; Roddenberry Center for Stem Cell Biology and Medicine at Gladstone, San Francisco, CA, USA; Department of Pediatrics, Cardiovascular Research Institute, Institute for Human Genetics, and Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94158, USA. Electronic address:
As light sheet fluorescence microscopy (LSFM) becomes widely available, reconstruction of time-lapse imaging will further our understanding of complex biological processes at cellular resolution. Here, we present a comprehensive workflow for in toto capture, processing, and analysis of multi-view LSFM experiments using the ex vivo mouse embryo as a model system of development. Our protocol describes imaging on a commercial LSFM instrument followed by computational analysis in discrete segments, using open-source software.
View Article and Find Full Text PDFMethods Mol Biol
December 2024
Department of Chromosome Science, National Institute of Genetics, Shizuoka, Japan.
The bipolar shape of the microtubule-based spindle is a pivotal morphological phenotype for accurate chromosome segregation during cell division. However, existing descriptions of spindle morphogenesis remain largely qualitative. Here, we introduce a method that provides a quantitative description of the morphological growth dynamics of spindles using Xenopus egg cytoplasmic extract and a computational image analysis pipeline.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
November 2024
Van der Waals-Zeeman Institute, Institute of Physics, University of Amsterdam, Amsterdam 1098XH, The Netherlands.
Photosynthetic algae play a significant role in oceanic carbon capture. However, their performance is constantly challenged by fluctuations in environmental light conditions. While phototaxis is a common strategy to cope with such fluctuations, nonmotile species must adopt alternative mechanisms to avoid light-induced damage.
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