ArticleMethods in molecular biology (Clifton, N.J.)2014
Isolating specific embryonic cells of the sea urchin by FACS.
Article in Methods in molecular biology (Clifton, N.J.), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 14 citations in OpenAlex.
- Single-cell RNA sequencing of theeLife · 2021Article
- Developmental single-cell transcriptomics in the Lytechinus variegatus sea urchin embryo.Development (Cambridge, England) · 2021Article
- Methodologies for Following EMT In Vivo at Single Cell Resolution.Methods in molecular biology (Clifton, N.J.) · 2021Review
- Autofluorescence mediated red spherulocyte sorting provides insights into the source of spinochromes in sea urchins.Scientific reports · 2020Article
- Trapping, tagging and tracking: Tools for the study of proteins during early development of the sea urchin.Methods in cell biology · 2019Review
- Differential Nanos 2 protein stability results in selective germ cell accumulation in the sea urchin.Developmental biology · 2016Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Isolating cells based on specific gene expression enables a focused biochemical and molecular analysis. While cultured cells and hematopoietic cells, for example, are routinely isolated by fluorescence activated cell sorting (FACS), early embryonic cells are a relatively untapped source for FACS applications often because the embryos of many animals are quite limiting. Furthermore, many applications require genetic model organisms in which cells can be labeled by fluorescent transgenes, or antibodies against cell surface antigens. Here we define conditions in the sea urchin embryo for isolation of embryonic cells based on expression of specific proteins. We use the sea urchin embryo for which a nearly unlimited supply of embryonic cells is available and demonstrate the conditions for separation of the embryo into single cells, fixation of the cells for antibody penetration into the cells, and conditions for FACS of a rare cell type in the embryo. This protocol may be adapted for analysis of mRNA, chromatin, protein, or carbohydrates and depends only on the probe availability for the cell of interest. We anticipate that this protocol will be broadly applicable to embryos of other species.
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