New look and touch screen compatible, but still three easy steps in the software to obtain concentrations:
- Image Processing: Acquire/import images and align a plate overlay.
- Well Assignment: Assign wells a type, name (optional), and dilution factor.
- Data Analysis: Customize and copy or export the automatically generated concentrations, statistics, and charts.
– You’ve just finished the last step of the assay:
- Capture the image(s): Acquire on a Q-View Imager or Import a grayscale 16-bit TIFF. New Image Processing produces lower %CVs at the low end of the curve and eliminates stacking.
- Input the Software Product Code: Found on the kit product card.
- Place the overlay: Use Image Options to optimize image gamma, zoom, and orientation for your convenience. Use the Overlay Options tools to align the plate overlay. Always finish with Auto-Adjust Spots. (Also Blot Tools)
- Assign Wells: Use Sequential Naming and Templates to quickly assign repeat layouts. Sample Controls can be assigned if they are part of the product definition.
- Analyze Data: Click Perform Analysis to fit using the new optimal regression settings or modify via dialog, default assay limits. Then choose Statistics & optimal dilution, mask outliers, compare curve fit and weighting options. Example use of new Help button. Copy or Export Charts or data out as needed.
Data Analysis Troubleshooting
If a curve or sample did not perform as expected, first ensure that all plate overlay spots are aligned, the image has white spots on a black background, and dilution factors have been assigned. Then check the actual data, not just visual cues in the image.
- Evaluate the Standard Curve
- Check the standard curve charts, statistics for outliers to mask. Also consider masking a standard curve point if: %CV > 20%, or %Backfit ≠ 80% – 120%.
- Check for signs of compression: Are there saturated points? Are there repeating pixel intensities?
- Evaluate Sample Signals
- Are the assay limits set to the ULOQ & LLOQ and does the signal fall on the linear part of the curve?
- Is there high well or Negative Control background? Toggle Negative Well Subtraction setting.
- Did sample controls, if present, pass?
- Re-optimize the Curve Fit if Needed: Compare various regression models, weighting, and masking options.