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Embryonic and induced pluripotent stem cell staining and sorting with the live-cell fluorescence imaging probe CDy1
- Source :
- Nature Protocols. 6:1044-1052
- Publication Year :
- 2011
- Publisher :
- Springer Science and Business Media LLC, 2011.
-
Abstract
- Detecting and isolating specific types of cells is crucial to understanding a variety of biological processes, including development, aging, regeneration and pathogenesis; this understanding, in turn, allows the use of cells for therapeutic purposes, for which stem cells have emerged recently as invaluable materials. The current methods of isolation and characterization of stem cells depend on cell morphology in culture or on immunostaining of specific markers. These methods are, however, time consuming and involve the use of antibodies that may often make the cells unsuitable for further study. We recently developed a fluorescent small molecule named CDy1 (compound of designation yellow 1) that selectively stains live embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). This protocol describes detailed procedures for staining ESC and iPSC in live conditions and for fluorescence-activated cell sorting (FACS) of ESC using CDy1. Cell staining, image acquisition and FACS can be done within 6 h.
- Subjects :
- Anthracenes
Staining and Labeling
Morpholines
Regeneration (biology)
Induced Pluripotent Stem Cells
Cell Culture Techniques
Cell sorting
Biology
Flow Cytometry
Cell morphology
Molecular biology
Embryonic stem cell
General Biochemistry, Genetics and Molecular Biology
Cell biology
Cell culture
Stem cell
Induced pluripotent stem cell
Embryonic Stem Cells
Immunostaining
Fluorescent Dyes
Subjects
Details
- ISSN :
- 17502799 and 17542189
- Volume :
- 6
- Database :
- OpenAIRE
- Journal :
- Nature Protocols
- Accession number :
- edsair.doi.dedup.....7c1a5601db34503ba55624ed505f2229