title: Embryonic and induced pluripotent stem cell staining and sorting with the live-cell fluorescence imaging probe CDy1.
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.
authors: Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium, Agency for Science, Technology and Research (A*STAR), Singapore, Republic of Singapore.
authors: Nam-Young
authors: NY
authors: Kang
authors: Seong-Wook
authors: SW
authors: Yun
authors: Hyung-Ho
authors: HH
authors: Ha
authors: Sung-Jin
authors: SJ
authors: Park
authors: Young-Tae
authors: YT
authors: Chang
mesh_terms: D000873:Anthracenes / Q000032:analysis* / Q000737:chemistry; D018929:Cell Culture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D009025:Morpholines / Q000032:analysis* / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000873:Anthracenes; C568650:CDy1 compound; D005456:Fluorescent Dyes; D009025:Morpholines