title: Imaging Pluripotency: Time-Lapse Analysis of Mouse Embryonic Stem Cells.
abstract: The current view of the pluripotent state is that of a transient, dynamic state, maintained by the balance between opposing cues. Understanding how this dynamic state is established in pluripotent cells and how it relates to gene expression is essential to obtain a more detailed description of the pluripotent state.In this chapter, we describe how to study the dynamic expression of a core pluripotency gene regulator-Nanog-by exploiting single-cell time-lapse imaging of a reporter mESC line grown in different cell culture media. We further describe an automated image analysis method and discuss how to extract information from the generated quantitative time-course data.
authors.affiliation: Faculdade de Medicina da Universidade de Lisboa, Instituto de Medicina Molecular and Instituto de Histologia e Biologia do Desenvolvimento, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.
authors.forename: Anna
authors.initials: A
authors.lastname: Pezzarossa
authors.affiliation: Faculdade de Medicina da Universidade de Lisboa, Instituto de Medicina Molecular and Instituto de Histologia e Biologia do Desenvolvimento, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.|Champalimaud Neuroscience Programme, Champalimaud Centre for the Unknown, Avenida Brasilia-Doca de Pedrouços, 1400-038, Lisbon, Portugal.
authors.forename: Ana M V
authors.initials: AM
authors.lastname: Guedes
authors.affiliation: Faculdade de Medicina da Universidade de Lisboa, Instituto de Medicina Molecular and Instituto de Histologia e Biologia do Desenvolvimento, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.|Champalimaud Neuroscience Programme, Champalimaud Centre for the Unknown, Avenida Brasilia-Doca de Pedrouços, 1400-038, Lisbon, Portugal.
authors.forename: Domingos
authors.initials: D
authors.lastname: Henrique
authors.affiliation: Faculdade de Medicina da Universidade de Lisboa, Instituto de Medicina Molecular and Instituto de Histologia e Biologia do Desenvolvimento, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal. elsa.abranches@gmail.com.|Champalimaud Neuroscience Programme, Champalimaud Centre for the Unknown, Avenida Brasilia-Doca de Pedrouços, 1400-038, Lisbon, Portugal. elsa.abranches@gmail.com.
authors.forename: Elsa
authors.initials: E
authors.lastname: Abranches
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002453:Cell Cycle; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods*; D005786:Gene Expression Regulation; D018398:Homeodomain Proteins / Q000032:analysis* / Q000235:genetics; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D000071317:Nanog Homeobox Protein; D061848:Optical Imaging / Q000379:methods; D059010:Single-Cell Analysis / Q000379:methods*; D013997:Time Factors
chemical_list: D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse
keywords: Dynamics; Heterogeneity; Nanog; Pluripotency; Stem cells; Time-lapse imaging