Tips for Data Analysis in the Q-View™ Software

The Q-View Software is a tool for the quantitative analysis of multiplexed chemiluminescent or infrared fluorescent assays, such as those found in Q-Plex™ planar-based arrays. The software enables users to:

  • Aquire images of microarrays and stack images optimized for bright and dim reactions, obtaining a single high dynamic range image. 
  • Easily locate spots for each assay using the software’s Auto-Set Plate Overlay and Auto-Adjust Spots features.
  • Assign wells as samples, controls, standards, or negatives, and specify their dilution factors.
  • Fit curves using one of several curve-fitting models including 4 and 5 Parameter Logistic (PL), and customize or export the automatically generated charts and reports. 

How does Q-View calculate concentrations?

Once the plate overlay has been placed, foreground pixels within each overlay circle are averaged using a proprietary weighting algorithm which is designed to prevent well background from having a large impact on the calculation.

How do I know what curve fitting model to choose?

We recommend the 4PL or 5PL regression models for Q-Plex data, as these are generally considered to provide the best overall fit for immunoassay curves. Use the Auto-Select option to have the Q-View Software automatically fit each curve using the model with the lowest AIC statistic. If your data is abnormal or you are trying to optimize the fit in a particular range (see below), one of the other Q-View regression models may better fit your needs.

When should I mask a standard curve point?

Consider masking a standard curve point if the %CV between the standard curve replicates is greater than 20%, or if the average %Backfit is outside 80%-120%. You might also consider masking if you are trying to optimize the fit in a particular range (see below).

Can I optimize the curve fit for the low or high end of the curve?

Yes, in the Data Analysis window navigate to the Curve Fit Options menu > Show Advanced Options to reveal different weighting priorities. The Software Product Code will default to the settings optimized by our own QC process but if your samples require it, try selecting “Auto Select Favor Low” or “Auto Select Favor High” to better fit your data.  

Previous Immunoassay Signal and Evaluating Standard Curves