Gating strategiesThe purpose of seq

Gating strategiesThe purpose of sequential gating is to select the population of interest and simultaneously minimize interference from debris and mature cells to which antibodies can bind nonspecifically. A gating strategy based on CD45 staining versus side-angle light scatter properties is suggested as a primary gate due to its utility in separating WBC from red cells, platelets, and other debris. Although nucleated red cells and fully mature plasma cells are not detectably stained by CD45-FITC, these cells are not generally found in significant numbers in PBSC preparations and so do not affect the utility of CD45+ events to generate a stable denominator in the calculation of the absolute CD34+ value. From this primary gate, CD34+ cells are identified by their expression of CD34, their characteristic low to intermediate CD45 antigen expression, and their characteristic light scatter properties.