Figure 9

Automation of Qdot-RPPA Image and Data Analysis. Several features of MicroVigene™ software are to facilitate the automatic batch process. A. Super Curve: Instead of generating multiple linear regression curves for each serial dilution, the Super Curve algorithm uses all spots on the same array to fit one antigen-antibody reaction curve (called Super Curve, the red line) for the quantitative readouts. The purple lines indicate error boundary. B. 2D and 3D views of the actual spot boundary algorithm for actual spot identification and dust finding (indicated by asterisk); the total intensity within the defined spot boundary (i.e. volume) was used for Super Curve fitting. C. Screen shots of the dust removal feature. Left: before removing the dust, the spot is flagged as an outlier (the red spot pointed by the arrow). Right: after dust removal, the spot fits the Super Curve.