2.6 Flow cytometry
Neutrophils were analysed by flow cytometry to quantify the percentage
of neutrophils: a) remaining morphologically intact, b) showing markers
of apoptosis, c) containing myeloperoxidase, after 18 h of adhesion to
fibrinogen coated surfaces. To this end, neutrophils were detached from
fibrinogen by brief exposure to EDTA/EGTA (both 5 mMol/L). Neutrophil
apoptosis was quantified by a FITC-conjugated Annexin-V Kit according to
the manufacturer’s instruction (BD-Biosciences, USA). Intracellular
myeloperoxidase content was measured by the ”MPO-FITC” Kit according to
manufacturer instruction (Beckman Coulter, Milan, Italy).