Metaxpress high content image acquisition analysis software

For all experiments, unless otherwise indicated, the ImageXpress Micro XLS Widefield High-Content Analysis System (Molecular Devices^®, San Jose, CA, USA) was used to image cells. The images were analyzed using the applicable modules of the MetaXpress^® High-Content Image Acquisition & Analysis Software supplied by Molecular Devices^® (San Jose, CA, USA).

Glass-bottom 96-well plates (Greiner Bio-One, Morone, NC) were filled with 1 M NaOH for 1 h, washed several times with ultra-pure distilled water and then dried by a stream of nitrogen gas. Planar lipid bilayers decorated with scFv ligands were assembled in wells. Jurkat T cells were stained with 5 µM calcein-acetoxymethyl ester (AM) (Sigma-Aldrich, St. Louis, MO, USA) for 45 min at 37°C. 1 × 10⁵ cells were added per well and incubated at 37°C for 30 min. Unbound cells were removed by serial washes with PBS. The number of bound cells was measured at four sites per well using an ImageXpress Micro XL High-Content Screening System (Molecular Devices, CA, USA). Counts from eight sites (two wells) per each group were analyzed using MetaXpress High-Content Image Acquisition & Analysis Software (Molecular Devices, CA, USA).