Hi, I am working on the phenotypic screening of antiviral molecules. As a validation test for hits, it was suggested that I use the ‘Strictly Standardised Mean Difference’ (SSMD) analysis method, which does indeed appear to be more robust than a t-test or a Z-score. I perform the calculations using 96-well plates containing uninfected negative controls and infected controls. The first SSMD I calculate is the one that looks at the difference between the molecules and the infected positive controls. I consider a hit to be a molecule with an SSMD < -2. I then calculate another SSMD, this time looking at the difference between the molecules and the negative controls based on cell count. A good hit must have an SSMD greater than -1; otherwise, this indicates that it has cytotoxic properties. I then plot these two SSMD values on two axes and, using the thresholds given above, define a region where the hits are located for an experiment carried out in a 96-well plate. My question: I would like to group the results of several experiments onto a single graph. Is it necessary to include the negative and positive controls on the graph, given that the SSMD values obtained for the molecules are already normalised relative to these same controls? This question arises because I am unable to find any recent articles on this method, which seems to be little used yet is nevertheless interesting. Could you provide me with any recent references if you have any? Thanks François