10-color Panel Design for B-cell Screening
Dr. Joseph DiGiuseppe
Tuesday, May 29, 2018
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Chapters
Introduction
Standardization in clinical flow cytometry: The EuroFlow™ Consortium
EuroFlow Lymphoid Screening Tube (LST)
Standardization in clinical flow cytometry: The 2006 Bethesda Consensus conference
Bethesda conference: Consensus reagents for the initial evaluation of B-cell neoplasia
Existing 8-color screening tube for B-cell neoplasia (HH)
BD FACSCanto™ 10-color system permits incorporation of two additional markers, but which two?
Panel optimization: Know your instrument.
Relative fluorochrome strength: CD4
Spillover/spread analysis: BV421
Spillover/spread analysis: PerCP-Cy5.5
Spillover/spread analysis: APC
Summary of spillover/spread analyses
Panel design is an iterative process.
Where to put CD22?
CD22-APC-R700 conjugate: Normal marrow
More options for CD22
Panel design is an iterative process.
Moving immunoglobulin light chains?
Spread implications of Kappa/Lambda-BV605
Are mouse monoclonals the best anti-light-chain reagents?
Feasibility of combination: Normal marrow
Normal marrow gated on B cells
What about the remaining channel? Fluorescence minus one (FMO)
One last (slight) modification…
Abnormal cases: CLL
B lymphoblastic leukemia
Biclonal B-cell LPD
Relapsed B-LL s/p blinatumomab
Introduction
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