After 1 h incubation at 37°C, unbound cells were removed, and fraction of bound cells was determined by a fluorescence plate reader (Cyto Fluor II; PerSeptive Biosystems, Framigham, MA, USA) (excitation, 485 nm, emission, 530 nm).
Media and planktonic cells were removed.
Therewith the non-adhering cells were removed.
After 1 hr, floating cells were removed.
Nonadherent cells were removed.
Non-adherent cells were removed.
After washing, bound cells were recovered by removing the aluminum samples to LB+Cm/Glu and growing at 37 °C with shaking overnight.
The columns were removed from the magnetic field and bound cells were eluted by allowing 4 ml of FACS buffer to pass through the column by gravity at 4°C.
Nonadherent monocytes were removed by gentle washing with PBS and the bound cells were measured by a fluorescence plate reader at excitation/emission wavelengths of 485/535 nm (Millipore, Milford, MA)[27].
The column was washed, removed from the magnet and bound cells were washed out.
