Fig. 1.

Single-cell antibody nanowell process. Arrays of nanowells with dimensions of 50 μm × 50 μm × 50 μm were used for microengraving. Peripheral blood mononuclear cells were loaded into the nanowells. Cells in the nanowells were imaged using an automated epifluorescence microscope. Micrograving was performed by hybridizing nanowells with capture slides containing antihuman immunoglobulins for 1 h at 37 °C with 5 % CO₂. After incubation, nanowells containing intact live cells and capture slides were separated. A mixture of goat antihuman immunoglobulin G (IgG)-Alexa Fluor 647 (AF647) and fluorochrome-conjugated SSA/Ro60-AF488 and SSB/La-AF550 were added to the capture slides. Micrographs of microarrays were generating by scanning using a GenePix Autoloader 4200AL microarray scanner. The schematic has been modified from a previous study [17]