Cells can be sorted as a preparitive technique for a number of assays, including molecular, functional, and in vivo assays.
This example from Dr. Anne Galy's work shows how a pure hematopoietic progenitor cell population is produced by cell sorting. The top panel shows the "pre sort" immunophenotype of the sample. First, a gating region is drawn around the viable (PI-) lineage negative (Lin-) population (top left plot). Since no membrane permeabilization step was taken, the propidium iodide (PI) acts as a viablity stain not unlike trypan blue: PI- cells are "live" and PI+ cells are "dead". The lineage parameter consists of a cocktail (mixture) of many mature markers (e.g., CD3 for T cells) all conjugated to the same fluorochorme (FITC). Immature cells are negative for these markers and are said to be "lineage negative". The rest of the plots show only these gated events. For sorting, more gating regions are used to select PI-Lin-CD10-CD34+CD38+ progenitor cells. The bottom panel shows the very pure (up to 99%) "post sort" immunophenotype.
Wesa AK, Galy A. IL-1 beta induces dendritic cells to produce IL-12. Int Immunol. 13(8):1053-61, 2001.
Christopherson I, Piechoki M, Liu G, Ratner S, Galy A. Regulation of L-selectin expression by a dominant negative Ikaros protein. J Leukoc Biol. 69(4):675-83, 2001.
Wesa AK, Galy A. Regulation of T cell cytokine production by dendritic cells generated in vitro from hematopoietic progenitor cells. Cell Immunol. 208(2):115-24, 2001.
Yan Y, Panos JC, McCormick DJ, Wan Q, Giraldo AA, Brusic V, David CS, Kong YM. Characterization of a novel H2A-E+ transgenic model susceptible to heterologous but not self thyroglobulin in autoimmune thyroiditis: Thyroiditis transfer with Vß8+ T cells. Cellular Immunolology 212:63-70, 2001.
Ferlazzo G, Klein J, Paliard X, Wei WZ, Galy A. Dendritic cells generated from CD34+ progenitor cells with flt3 ligand, c-kit ligand, GM-CSF, IL-4, and TNF-alpha are functional antigen-presenting cells resembling mature monocyte-derived dendritic cells. J Immunother. 23(1):48-58, 2000.
Ferlazzo G, Wesa A, Wei WZ, Galy A Dendritic cells generated either from CD34+ progenitor cells or from monocytes differ in their ability to activate antigen-specific CD8+ T cells. J Immunol. 163(7):3597-604, 1999.
Cancovic M. CD4+ lymphocytes as a target for cyclophosphamide therapy of SJL lymphomas. PhD thesis. 1995.