Figure 4.

Isolation of cells by the OAC and sort-purification methods result in similar phagocytic function. Neutrophils were isolated from 5-month-old B6.Faslpr/J mice using the OAC protocol or by sort-purification. Isolated neutrophils were incubated in the presence of fluorescently labeled S. aureus particles with prior opsonization for 2 h. Engulfment of opsonized S. aureus by neutrophils was confirmed by high-resolution microscopy and flow cytometry using ImageStreamX technology. One representative image from more than 250 images taken from two separate animals for each purification method is shown. Uptake of S. aureus for each neutrophil was calculated as an internalization score (I.S.). The I.S. from 50 randomly selected neutrophils for each isolation method is shown in the right panel, demonstrating that the uptake of particles by neutrophils is equivalent between the two isolation methods.