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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.

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Presentation on theme: "Analysis and Sorting of T reg Cells on the FACSAria John Daley Director of Flow Cytometry Hematologic Neoplasia Dana-Farber Cancer Institute Harvard Medical."— Presentation transcript:

1 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 http://www.bdbiosciences.com/features/products Picture credit:

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

3 The way we used to sort Human CD4+CD25+ PBL’s on the FACSAria SORT TOP 2- 3% CD25 HI

4 Then CD45Ra was added to the Sort Design

5 Four way reg sort Multicolor(5) Minor populations : Four way sorting: Treg Story CD45ra FITC CD127 PE CD25 PE-CY5 CD4 PE-CY7 CD3 PAC BLUE

6 Compensation Comparisons Manual Comp with cells and actual reagents Autocomp with cells and actual reagents Autocomp with beads and actual reagents Autocomp with beads and CD45+ reagents Autocomp with cells and CD45+ reagents

7 BD cells Cell Autocomp BD cells Bead AutoComp

8 Treg Post Sort Reanalysis

9 99.497.1 98.4 99.9 POST SORT REANALYSIS: HUMAN PBL CD3+CD4+ CD45RA+ &CD45RA- Treg+/Teffector+/ --

10 Actual reagents cells compensation cells beads Actual reagents beads compensation cells beads

11 FoxP3 Expression in CD4+ Treg CD45RA+

12 Fluorescence minus 1

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