ArticleJournal of visualized experiments : JoVE2016
Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos.
Article in Journal of visualized experiments : JoVE, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Systematic evaluation of computational methods for cell segmentation.Briefings in bioinformatics · 2026Article
- ZFP281 controls transcriptional and epigenetic changes promoting mouse pluripotent state transitions via DNMT3 and TET1.Developmental cell · 2024Article
- Adaptative response to changes in pyruvate metabolism on the epigenetic landscapes and transcriptomics of bovine embryos.Scientific reports · 2023Article
- Enhancer-promoter interactions can bypass CTCF-mediated boundaries and contribute to phenotypic robustness.Nature genetics · 2023Article
- Article
- RNA Pol II pausing facilitates phased pluripotency transitions by buffering transcription.Genes & development · 2022Article
- Extensive co-binding and rapid redistribution of NANOG and GATA6 during emergence of divergent lineages.Nature communications · 2022Article
- Klf4 methylated by Prmt1 restrains the commitment of primitive endoderm.Nucleic acids research · 2022Article
- Simultaneous visualization of RNA transcripts and proteins in whole-mount mouse preimplantation embryos using single-molecule fluorescenceFrontiers in cell and developmental biology · 2022Article
- Live Visualization of ERK Activity in the Mouse Blastocyst Reveals Lineage-Specific Signaling Dynamics.Developmental cell · 2020Article
- Article
- 3D-Cell-Annotator: an open-source active surface tool for single-cell segmentation in 3D microscopy images.Bioinformatics (Oxford, England) · 2020Article
- Software tools for 3D nuclei segmentation and quantitative analysis in multicellular aggregates.Computational and structural biotechnology journal · 2020Review
- Article
- ABiology open · 2018Article
- A Sprouty4 reporter to monitor FGF/ERK signaling activity in ESCs and mice.Developmental biology · 2018Article
- Article
- Article
- Quantitative Whole-mount Immunofluorescence Analysis of Cardiac Progenitor Populations in Mouse Embryos.Journal of visualized experiments : JoVE · 2017Article
- Lineage Establishment and Progression within the Inner Cell Mass of the Mouse Blastocyst Requires FGFR1 and FGFR2.Developmental cell · 2017Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
This protocol presents a method to perform quantitative, single-cell in situ analyses of protein expression to study lineage specification in mouse preimplantation embryos. The procedures necessary for embryo collection, immunofluorescence, imaging on a confocal microscope, and image segmentation and analysis are described. This method allows quantitation of the expression of multiple nuclear markers and the spatial (XYZ) coordinates of all cells in the embryo. It takes advantage of MINS, an image segmentation software tool specifically developed for the analysis of confocal images of preimplantation embryos and embryonic stem cell (ESC) colonies. MINS carries out unsupervised nuclear segmentation across the X, Y and Z dimensions, and produces information on cell position in three-dimensional space, as well as nuclear fluorescence levels for all channels with minimal user input. While this protocol has been optimized for the analysis of images of preimplantation stage mouse embryos, it can easily be adapted to the analysis of any other samples exhibiting a good signal-to-noise ratio and where high nuclear density poses a hurdle to image segmentation (e.g., expression analysis of embryonic stem cell (ESC) colonies, differentiating cells in culture, embryos of other species or stages, etc.).
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Registered trials
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