Differentiating between fertilized and unfertilized eggs

I am studying fertilization success in sea urchins and I am looking for a way to count fertilized versus unfertilized eggs from pictures. The biggest difference between the two is circularity, as the fertilized eggs have visible bumps around the edges. I also need to be able to differentiate between cells when they are close together, as the other program I have tried using (ImageJ) counts eggs near each other as one object. I have attached an image so you can see what I am working with. Right now there are not many eggs per picture because I am counting them by hand, but I would like to find a way to take one picture of each well and count it all at once.
Thanks,
Tess


Hi Tess,

Sorry for the delay in response. I’ve put together a pipeline that might help. A few comments:

  • I split the color image into its component parts so I could pick out the color channel that had the best contrast, in this case, the red channel.
  • I tried to remove the well exterior from consideration, but this approach may lead to erroneous results if the well is *not *
    visible in the picture. - Then some illumination correction to remove the spatial heterogeneities in the background intensity.
  • After identifying the eggs, it seemed that solidity was a good measure to filter by; according to the help, solidity is the “proportion of the pixels in the convex hull that are also in the object, i.e. ObjectArea/ConvexHullArea. Equals 1 for a solid object (i.e., one with no holes or has a concave boundary), or <1 for an object with holes or possessing a convex/irregular boundary.” So an object with indentations will have a solidity somewhat less than 1. The DisplayDataOnImage module just beforehand is a good way to preview the values of a given measurement on the image itself.

Good luck!
-Mark
pipeline.cppipe (13.2 KB)

Thank you very much for your help!