C. albicans blastoconidia were allowed to elaborate into hyphae on tissue culture plastic coated with 10 μg/ml Fn in YPD at 37°C. Hyphae were washed and human neutrophils, pretreated on ice with 10−9 M fMLP, were added and incubated at 37°C in L-15 supplemented with 2 mg/ml glucose and 1 mM Mn++. After 30 min incubation, NET formation was visualized using Sytox green, imaged, and scored for cluster formation and NETs with (A) control cells and (B) neutrophils that were additionally pretreated either with specific mAbs to block VLA5 (P1D6) or VLA3 (MKID-2) or to activate VLA5 (SNAKA51). Bright field and FITC images acquired at 20x; bar = 100μm. (C) Enlarged images showing cluster aggregation to hyphae.