Human memory T cell purity analysis (pre- and post-FACS sort)

Human blood was subjected to Ficoll while enriching for CD4+ cells using Stem Cell Tech Rosette Sep. Cells were then stained with Fc block, followed by Alexa700-anti-human CD4, PE-anti-human CD45RO, APC-anti-human CD45RA, and a FITC dump panel (CD235a, CD56, CD36, CD19, CD16, CD14). Stained cells were sorted on a FACSAria. Conclusion: We have found in subsequent experiments that the 0.2-0.5% of cells outside the CD4+ gate are dying cells - purity is increased to greater than 99.9% when a subsequent post-sort stain with live/dead Aqua amine is performed and cells are reanalyzed gating on live/dead aqua amine negative cells. Notes: Cytobank experiment no. 518.