Sequential gates and the actual denominator

The gate order is acquired events (180,000), debris-excluded events (171,000), singlets (162,000), viable events (158,000), CD45-positive leukocytes (154,800), then the CD45-dim, low-side-scatter target (43,347). The target is 43,347 of 154,800 leukocytes: 28.00194%, not a rounded 28%. Low side scatter is an observed characteristic of this target gate, not an independent lineage assignment.

Positive means fluorescence above the locally established negative boundary in the stated gate. Negative means it does not exceed that boundary in the stated gate; neither word is an absolute biologic absence or presence. Bright and dim mean relatively higher and lower intensity than the method's relevant internal comparator. Uniform means the defined gated population has a narrow, single expression distribution; heterogeneous means it contains reproducible differing distributions. Coexpression means the same events are demonstrated positive for two markers on a paired-marker display in the same gate. Marginal percentages from separate markers or tubes never prove coexpression.

The figure contains every population count, each parent, the target count and unrounded percentage, and the low-side-scatter observation. Marker percentages belong to the target gate and must not be presented as a percentage of viable leukocytes.