Establishment of a whole human blood immobilized reporter assay. (A) View of the whole human blood flow chamber coated with the LFA-1 activation reporter antibodies. (B) Cells rolling in the microflow chambers visualized by transillumination. Scale bar = 100 μm. (C) Flow chambers coated with isotype control, TS1-22 and/or E-selectin were perfused with heparinized human whole blood for 2 minutes at 5.94 dyn/cm2, washed with HBSS with 1mM Ca2+ and 1mM Mg2+ for 30 seconds, fixed with 2% paraformaldehyde, and stained with Wright-Giemsa stain solution and number of neutrophils counted. Some flow chambers were incubated with E-selectin blocking antibody IBIG-E4 before perfusion, and number per field of view was calculated. Data are presented as mean ± SEM and are representative of 3 independent experiments.
