Net signal intensities and backgrounds. Serum samples were incubated on antibody microarrays prepared on hydrogels and nitrocellulose and detected with direct labeling, indirect detection, and RCA. Two different serum samples were measured in duplicate for each condition, using the same serum sample in both color channels (635 and 532). The net signal is the background-subtracted, median intensity of each antibody spot, averaged over the four replicate experiments. Scanner settings were identical within microarrays performed on the same substrate. (a-d) The distribution of average background-subtracted intensities of the antibody spots for the indicated substrates, color channels and detection methods. (e, f) The average background levels for each detection method and substrate.
Zhou et al. Genome Biology 2004 5:R28