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Sloan Kettering Institute

3 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos
Nestor Saiz 1, Minjung Kang 1, Nadine Schrode 1, Xinghua Lou 1, Anna-Katerina Hadjantonakis 1
1Developmental Biology Program, Sloan Kettering Institute

This protocol presents a method to perform quantitative, single-cell in situ analysis of protein expression to study lineage specification in mouse preimplantation embryos. The procedures necessary for collection of blastocysts, whole-mount immunofluorescent detection of proteins, imaging of samples on a confocal microscope, and nuclear segmentation and image analysis are described.

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Genetics

Genome Editing and Directed Differentiation of hPSCs for Interrogating Lineage Determinants in Human Pancreatic Development
Zhong-Dong Shi *1, Chew-Li Soh *1, Zengrong Zhu *1, Danwei Huangfu 1
1Developmental Biology Program, Sloan Kettering Institute

Protocols to generate hPSC mutant lines using the iCRISPR platform and to differentiate hPSCs into glucose-responsive β-like cells are described. Combining genome editing technology with hPSC-directed differentiation provides a powerful platform for the systematic analysis of the role of lineage determinants in human development and disease progression.

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Developmental Biology

Isotropic Light-Sheet Microscopy and Automated Cell Lineage Analyses to Catalogue Caenorhabditis elegans Embryogenesis with Subcellular Resolution
Leighton H. Duncan 1,5, Mark W. Moyle 1,5, Lin Shao 1,5, Titas Sengupta 1,5, Richard Ikegami 1,5, Abhishek Kumar 4,5, Min Guo 4,5, Ryan Christensen 4,5, Anthony Santella 2,5, Zhirong Bao 2,5, Hari Shroff 4,5, William Mohler 3,5, Daniel A. Colón-Ramos 1,5,6
1Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine, 2Developmental Biology Program, Sloan Kettering Institute, 3Department of Genetics and Genome Sciences and Center for Cell Analysis and Modeling, University of Connecticut Health Center, 4Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, 5WormGUIDES.org, 6Instituto de Neurobiología, Recinto de Ciencias Médicas, Universidad de Puerto Rico

Here, we present a combinatorial approach using high-resolution microscopy, computational tools, and single-cell labeling in living C. elegans embryos to understand single cell dynamics during neurodevelopment.

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