I have cells growing on an oriented substrate and I'm looking to validate the orientation module for my pipeline. I use the MeasureObjectSizeShape module in CellProfiler 2.1.1. To that end, I started by drawing identical circles in PowerPoint, then altering their dimensions to go systematically from a 1:1 ellipse to a 2:1 ellipse, all in the same orientation. All the shapes were saved as .tif files.
My problem comes with the circles (1:1 ellipses if you will). I was expecting a random distribution of orientations, but have instead obtained a distinct mode around -36 degrees. I'm taking this to be due to the pixellation that is part and parcel of any raster file.
My question is thus in two parts. First, is it possible to have vector graphics open in CellProfiler to verify that what I'm seeing is an artefact? Second, and more fundamentally, has anyone advice on what images are a good way to validate the software modules?
A loyal CellProfiler User