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Live Imaging of Mouse Embryonic Stem Cells and Embryos: Protocols for In Vitro Culture, Microscopy, and Image Data
Woonyung Hur1,2, Mohamed I Gatie1, Alexandre Francou1
1Developmental Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
Methods in Molecular Biology (Clifton, N.J.)
|December 28, 2025
Summary
This study provides protocols for live imaging of mouse pluripotent stem cells and embryos using advanced microscopy. These methods enable quantitative analysis of cell dynamics and developmental processes with minimal phototoxicity.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Microscopy
Background:
- Live imaging is crucial for observing dynamic cellular processes in developmental and stem cell biology.
- Current protocols require optimization for long-term, high-resolution imaging of sensitive biological samples.
Purpose of the Study:
- To present optimized protocols for live imaging of mouse pluripotent stem cells (mPSCs) and embryos.
- To enable quantitative analysis of cell state transitions and morphogenetic processes.
Main Methods:
- Utilizing confocal and light-sheet microscopy for live imaging.
- Implementing optimized ex utero culture, mounting, and environmental control systems.
- Employing computational workflows for image processing, segmentation, and cell tracking.
Main Results:
- Established protocols support long-term, physiologically relevant live imaging of mPSCs and embryos.
- Minimized phototoxicity during imaging sessions.
- Enabled quantitative lineage dynamics analysis.
Conclusions:
- The presented protocols offer a comprehensive and reproducible framework for studying mammalian development.
- Facilitates investigation into cell state transitions and morphogenesis using live imaging techniques.
