Counting beads in droplets

Hello and thank you for this amazing tool! I am very new to image analysis and I am working with droplets that contain beads. I want to answer to 2 questions:

  1. What is the size of my droplets and the SD
  2. How many beads I have per droplet

I am using this pipeline:
droplet analysis.cpproj (415.7 KB)

and I have this kind of pictures:Image 000028.tif (2.7 MB)

So I can identify droplets and have the information about size and SD, but I don’t understand how to identify and count the beads. Can you please help me with this?

Thank you!


Hi @FrancescaB,

I checked your images & pipeline.

Here is my answer to your queries,

  1. Size of droplets: You had already identified them & have the info.
  2. Identifying beads: I have added few more modules in your pipeline towards this. Firstly, mask the region where you have the droplets (“MaskImage”) followed by inverting the image (“ImageMath”) and rescaled the intensity in order to make the thresholding easier (“RescaleIntensity”) & used this image to identify beads (with the same module settings you had used i.e.“IdentifyPrimaryObject”). Now you can count the beads from the image similar to how you counted the droplets.
  3. Number of beads on every droplets: In order get the number of beads from every droplet, you may have to use “RelateObjects” module to get the count in every droplet.
    Here is the screenshot of beads identification.
    Note: PFA pipeline. You may require some optimization in “IndentifyPrimaryObject” please check.

droplet analysis_LB.cpproj (404.5 KB)

Hope this helps.


Hi Lakshmi,

Thank you very much for your answer, it helped a lot!



1 Like