Sloan Kettering Institute 3 articles published in JoVE Developmental Biology Isotropic Light-Sheet Microscopy and Automated Cell Lineage Analyses to Catalogue Caenorhabditis elegans Embryogenesis with Subcellular Resolution Leighton H. Duncan1,5, Mark W. Moyle1,5, Lin Shao1,5, Titas Sengupta1,5, Richard Ikegami1,5, Abhishek Kumar4,5, Min Guo4,5, Ryan Christensen4,5, Anthony Santella2,5, Zhirong Bao2,5, Hari Shroff4,5, William Mohler3,5, Daniel A. Colón-Ramos1,5,6 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. Developmental Biology Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos Nestor Saiz1, Minjung Kang1, Nadine Schrode1, Xinghua Lou1, Anna-Katerina Hadjantonakis1 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. Neuroscience Feeder-free Derivation of Neural Crest Progenitor Cells from Human Pluripotent Stem Cells Nadja Zeltner1, Fabien G. Lafaille2, Faranak Fattahi1, Lorenz Studer1 1Developmental Biology, Center for Stem Cell Biology, Sloan-Kettering Institute for Cancer Research, 2St. Giles Laboratory of Human Genetics of Infectious Diseases, The Rockefeller University Neural crest (NC) cells derived from human pluripotent stem cells (hPSC) have great potential for modeling human development and disease and for cell replacement therapies. Here, a feeder-free adaptation of the currently widely used in vitro differentiation protocol for the derivation of NC cells from hPSCs is presented.
