Analysis and Sorting of T reg Cells on the FACSAria

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    08-Feb-2016
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Analysis and Sorting of T reg Cells on the FACSAria. John Daley Director of Flow Cytometry Hematologic Neoplasia Dana-Farber Cancer Institute Harvard Medical School. Picture credit:. http://www.bdbiosciences.com/features/products. From BD Biosciences Website Featured products: - PowerPoint PPT Presentation

Transcript of Analysis and Sorting of T reg Cells on the FACSAria

  • Analysis and Sorting of T reg Cells on the FACSAriaJohn DaleyDirector of Flow CytometryHematologic NeoplasiaDana-Farber Cancer InstituteHarvard Medical Schoolhttp://www.bdbiosciences.com/features/productsPicture credit:

  • http://www.bdbiosciences.com/features/products/display_product.php?keyID=162From BD Biosciences WebsiteFeatured products:Regulatory T cells

  • The way we used to sort Human CD4+CD25+ PBLs on the FACSAriaSORT TOP2- 3% CD25 HI

  • Then CD45Ra was added to the Sort Design

  • Multicolor(5) Minor populations : Four way sorting: Treg StoryCD45ra FITCCD127 PECD25 PE-CY5CD4 PE-CY7CD3 PAC BLUE

  • Compensation ComparisonsManual Comp with cells and actual reagentsAutocomp with cells and actual reagentsAutocomp with beads and actual reagentsAutocomp with beads and CD45+ reagentsAutocomp with cells and CD45+ reagents

  • BD cells Cell AutocompBD cellsBead AutoComp

  • Treg Post Sort Reanalysis

  • 99.497.198.4

    99.9POST SORT REANALYSIS: HUMAN PBL CD3+CD4+ CD45RA+ &CD45RA- Treg+/Teffector+/ --

  • Actual reagents cells compensationcellsbeadsActual reagents beads compensationcellsbeads

  • FoxP3 Expression in CD4+ Treg CD45RA+

  • Fluorescence minus 1