DURAClone IM Phenotyping Basic Principle of the Panel Design

Markers

 Specificity  Purpose Clone Fluorochrome Laser Filter
 CD45 Leukocyte gating  J33 Krome Orange 405nm 525/40
 CD3 T cell lineage UCHT1 APC-Alexa Flour 750 638 nm 763/43*
 CD4 T Helper cell lineage 13B8.2 APC 638 nm 660/10
 CD8 Cytotoxic T cell lineage B9.11 Alexa Fluor 700 638 nm 712/25
 CD19 B cell lineage  J3-119 ECD (PE-Texas Red) 488 nm 610/20
 CD14 Monocyte lineage  RM052 PE-Cy7 488 nm 763/43*
 CD16  NK cell lineage  3G8 FITC 488 nm 525/40
 CD56  NK cell lineage  N901 PE 488 nm 585/42

*Also detectable in 780/60 bandpass used in the CytoFlex and several of the CytoFLEX S series instruments.

Co-expression Patterns and Spillover Rules

DURAClone IM Phenotyping Basic Panel Design Rules

Hierarchical tree structure showing parent-child relationships between the interrogated antigens. Child nodes inherit the properties of the parent nodes above it in the tree.  To illustrate, all populations in the analysis will be CD45+, inheriting that property from the parent node. As a principle, spillover from parent to child nodes should be minimized, while spillover from child to parent nodes will not cause a loss of resolution.  Nodes at the same hierarchy level without dashed connections are co-expressed antigens whose resolution is impacted by spillover. Colored boxes indicate the expression pattern for the given marker, the lower the expression the more important it will be to avoid spillover from other dyes into that channel. Mutually exclusive antigens in the same level are indicated with a dashed red line.  Spillover between these detectors will not cause a loss of resolution as the signals will not be present on the same event.  Pairs of antigens that are either double-positive or double-negative are positively correlated antigens that can tolerate spillover, indicated with a dashed gold line.

 

For Research Use Only. Not for use in diagnostic procedures.