So… you might need to do a little pre-processing before any detection methods/tools. You have variation in background, etc. I am not sure this is the best (in fact, there are most likely better methods) - but I just did the following:
- Convert your image to 8-bit.
- Duplicate it and call the duplicate image “background”
- Apply a Gaussian blur (sigma = 250) the “background” image
- Use the Image Calculator and subtract the “background” image from the 8-bit original (create a 32-bit float).
Then you can try using Trackmate for particle detection. I know this is a tracking tool - but it’s object detection is awesome!
This is what I got for a first-go of your image after doing the above background adjustments (I used an estimated blob diameter 30 pixels):
So this could be an option for a workflow. Though perhaps the background method I used is not ideal - maybe someone else here on the forum has better insight on cleaning up your image a bit before detection?