1 / 12

Analysis and Sorting of T reg Cells on the FACSAria

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:

marina
Download Presentation

Analysis and Sorting of T reg Cells on the FACSAria

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

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

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

  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 Bead AutoComp BD cells Cell Autocomp

  8. Treg Post Sort Reanalysis

  9. POST SORT REANALYSIS: HUMAN PBL CD3+CD4+ CD45RA+ &CD45RA- Treg+/Teffector+/ -- 98.4 99.4 97.1 99.9

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

  11. FoxP3 Expression in CD4+ Treg CD45RA+

  12. Fluorescence minus 1

More Related