Multiplexing Phenotype and Function for More Biologically Relevant Insights - 17

More Biologically Relevant Insights | APPLICATION NOTE

Data from an example assay using Jurkat target cells and NK-92 effector cells is
shown in Figure 3. Figure 3A shows two distinct cell populations, the background
fluorescence of unlabeled NK-92 cells and a higher intensity signal generated by
fluorescence-encoded Jurkat cells.
Gating the Jurkat cells and analyzing just this population in the FL1 channel for
caspase 3/7 activation enables quantification of Jurkat apoptosis. Because only
apoptotic cells with activated caspase 3/7 will display fluorescence in the FL1 detec-

Figure 3. Analysis of iQue Screener CMC assay. Target cells are encoded
with a dye that fluoresces in the FL4 channel, enabling differentiation
from unstained effector cells (A). Both target and effector cells can then
be queried for viability (B, D) or caspase activation (C, E) separately,
even though both are present in the same well.

17

| January, 2019

tion channel, we can determine the percent of apoptotic target cells in the well by
comparing the number of FL1-positive cells (Figure 3B, right peak) to FL1-negative
cells (Figure 3B, left peak).
Similarly, by analyzing the data for the Jurkat target cells in the FL3 channel, we can
measure the relative amount of cell death mediated by the effector cells (Figure 3C).
The higher intensity peak consists of FL3-positive cells with damaged membranes
(Figure 3C, right peak). A gate can be drawn to quantify the number of target cells


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Multiplexing Phenotype and Function for More Biologically Relevant Insights

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