Abstract
This chapter details 3D morphological topography of colony architecture optimization and nuclear protein localization by co-immunofluorescent confocal microscopy analysis. Colocalization assessment of nuclear and cytoplasmic cell regions is detailed to demonstrate nuclear and cytoplasmic localization in mEpiSCs by confocal microscopy and orthogonal colocalization assessment. Protein colocalization within mESCs, mEpiLCs, and mEpiSCs can be efficiently completed using these optimized protocols.
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Acknowledgments
mESC and mEpiSC pluripotent stem cell lines were graciously gifted from Dr. Janet Rossant. This research was funded by a Canadian Institutes of Health Research operating grants to A.J.W. and D.H.B. and Natural Sciences and Engineering Research Council of Canada grant to D.H.B. The funding sources played no role in design, data collection, analysis, decision to publish, or preparation of this study and manuscript.
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Dierolf, J.G., Watson, A.J., Betts, D.H. (2022). 3D Immunofluorescent Image Colocalization Quantification in Mouse Epiblast Stem Cells. In: Osteil, P. (eds) Epiblast Stem Cells. Methods in Molecular Biology, vol 2490. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2281-0_7
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DOI: https://doi.org/10.1007/978-1-0716-2281-0_7
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