Figure 2: Examples of gating strategies for basophils:
  1. Basophils were identified as SSClow CD123c+ CD193+ cells: 1. Lymphocyte – monocyte gate on a FSC/SSC plot, 2. Doublet exclusion FSC-H vs FSC-A, then SSC-H vs SSC-H, 3. Gate on both markers simultaneously CD203c x CD193, 4. Set CD63 negative threshold to 2.5 % (was 5%, but is more universal at 2.5%), 5. Assess the positive population.
  2. Basophils were identified as SSClow CD203c+ CD123+ HLA-DR-4. The CD63 gate is set on the negative control and basophil activation is measured above the gate for the other stimulation conditions, either with allergen or positive controls.